>> MY NAME IS ANTONELLO PILEGGI AND I AM A PROGRAM OFFICER FROM THE NATIONAL INSTITUTE FOR CHILD AND HUMAN DEVELOPMENT. I WANT TO THANK YOU FOR JOINING US TODAY AND WE HAVE A FANTASTIC THURSDAY OF THIS WORKSHOP. YESTERDAY WE HAD A GREAT HEAD START WITH A VERY FORMATIVE PRESENTATION. WE COVERED DIFFERENT THINGS FROM REGULATORY MANUFACTURING TO EXAMPLES OF LESSONS LEARNED IN THE FIELD ATTRIBUTED BY YOUR PAINTING AND CONCERTS AS WELL AS THE EVOLUTION OF THE CHILD MEDICINE FIELD. THIS IS MY DESIGN AND I REALLY WANTED TO HAVE PERSPECTIVE FROM DIFFERENT ANGLES AND TRYING TO FOSTER SOME DIFFERENT I THINK IT IS THE SAME THING THAT WE ARE STARTING TODAY WITH A CONTINUATION OF THE PROGRAM THAT CLEARLY COVERS THIS AND CONTINUES TO ILLUMINATE US ON THE STATE OF THE ART OF AND TO ENHANCE THE NUMBER OF THINGS THAT WILL MAKE IT TO IMPACT THIS IN THE STANDARD OF CARE FOR PATIENTS. ESPECIALLY BECAUSE WE ARE A PHD FOR AND FOSTER GENETICS AND ECOLOGY OF THIS POPULATION. TODAY WE HAVE A NUMBER OF AND A CONVERSATION WITH THE KEYNOTE PRESENTATION AND WE WILL BE LATER TALKING ABOUT AND WE HAVE CLOSED TODAY. TRYING TO CONTINUE ON THIS COURSE YESTERDAY THE DIALOGUE ON THE DIFFERENT TASKS AND WE CAN PICTURE THE SCREEN JUST TO GET SOME HOUSEKEEPING ASPECTS WHILE WE ENSURE THAT OUR FAMILY IS JOINING US IN THE YOU CAN SEE MY SLIDES WE HAVE SOMETHING THAT I WANT TO STRESS IS WE HAVE AN ACTIVE IDEA SCALE CAMPAIGN WHICH IS INTENDED TO SOLICIT ENGAGEMENTS BY ALL STAKEHOLDERS. WE REALLY WOULD LIKE TO HEAR YOUR VOICES PLEASE AND SHARE YOUR IDEAS IN THIS ROOM, AND WHAT YOU THINK THE MOST SIGNIFICANT ADVANCES IN THE FIELD AND IN YOUR HOME FIELD AND WE CAN APPRECIATE THIS AND THIS PHASE OF STAKEHOLDERS THAT SPANS A LOT OF TERRITORY. THAT IS REALLY INTENDED AND REALLY WE WOULD LIKE TO HEAR FROM YOU WHAT DO YOU THINK OF THE MOST IMPORTANT ACCOMPLISHMENTS OF THE FIELDS ARE AND IN YOUR AREA OF EXPECT HER EXPERTISE WHERE THE FIELD IS GOING AND WHERE WE SHOULD BE AIMING AT THE FIELDS AND OF OBSTETRIC MEDICINE IN THE TREATMENT AND ITS THE IF YOUR FIELD IS NOT ADVANCED YET, WHAT YOU THINK ARE THE MAJOR TASKS AND HOW CAN WE THINK OF POSSIBLE SOLUTIONS TO THOSE? IN GETTING AND WHETHER THERE IS ANYTHING THAT WE NEED. YESTERDAY WE HEARD ONE REGULATORY PATHWAY IS BECOMING MORE ACCESSIBLE AND THEY ARE STILL CHALLENGES IN THIS AND ALSO LEARN ABOUT HOW CHALLENGING IS. [INAUDIBLE] THE MODELS TO INFORM THE SAFETY AND EFFICACY OF THE SOLUTION THAT APPLIED TO HUMANS. AND ALSO THINKING IN THIS SPECIFIC EVENT HERE WITH A GYNECOLOGICAL EVENT THINK OF THE PEDIATRICS. YOU HAVE DIFFERENT STAGES THAT GO FROM AGE ZERO TO ADULTHOOD. WE HAVE AND REQUIREMENTS. AS WELL AS IN THIS OB/GYN POPULATION WE HAVE CHALLENGES FOR THIS UNIQUE POPULATION. I WOULD LIKE TO REALLY SOLICIT YOUR INPUT AND BE ENGAGED. YOU CAN COMMENT ON THE IDEAS THAT OTHERS HAVE POSTED ON THE CAMPAIGN SO THAT WE CAN HAVE A VIABLE AND THIS WILL BE ACTIVE THROUGH DECEMBER 16th. REALLY THIS CAN HELP CONTINUE THE DIALOGUE AND ON THIS WORKSHOP. ALSO IT'S VERY IMPORTANT THIS ENGAGED INTERACTION WITH YOUR ATTENDEES AND PLEASE FEEL FREE TO SUBMIT YOUR QUESTIONS THROUGH THIS LIFE THE FEEDBACK IN THE VIDEO STREAM PAGE SO THE BOTTOM OF THE STREAM THESE WILL SEE THE LOGISTICS TEAM AND WILL SHARE IT SO THE PRESSING QUESTIONS THAT YOU HAVE CAN BE ASKED. YOU CAN ALSO PUT YOUR COMMENTS ON THIS SCALE PAINT CAMPAIGN THAT WILL GO BEYOND THIS MEETING TODAY. IF WE, LET'S SEE. IF WE HAVE OUR SPEAKERS HERE ARE NOT SURE IF I HAVE IT AND WANT TO SEE OUR KEYNOTE SPEAKER. OKAY. SO WELL THEN I THINK IF WE COULD START A LITTLE BIT EARLIER WE WOULD HAVE A CHANCE TO MAYBE GAIN MOMENTUM IN THIS DISCUSSION. LET'S SEE. TO INTRODUCE OUR KEYNOTE PRESENTER TODAY AND IN THIS COMMUNITY IS PROBABLY IS NOT INTERACTION BUT THE SURVEY EXCEPT IT'S VERY LONG CURRICULUM THIS VIDEO. CURRENTLY THE DIRECTOR OF THE WAKE FOREST INSTITUTE OF MEDICINE. HE IS A PROFESSOR AND ACTUALLY IS A PROFESSOR AND CHAIR OF THE DEPARTMENT AND THAT PER REFERENCE OF MOLECULAR MEDICINE AND DIRECTOR OF THE PROGRAM AND HE ALSO HAS. [INAUDIBLE] HE HAS A GREAT INTEREST THAT HAS BEEN A PIONEER AND MENTOR. HE HAS OBTAINED HIS BACHELOR OF THE UNIVERSITY OF MIAMI AND THEN HE HAS HIS MEDICAL DEGREE AND THE SURGICAL RELAY DIDN'T SEE AT THE UNIVERSITY IN KENTUCKY. AND THEN HE DID A RESEARCH FELLOWSHIP IN THE SCHOOL IN BOSTON FOR HIS CAREER SKYROCKETED AND HE HAS A NUMBER OF WORDS AND ACCOMPLISHMENTS HE IS RECEIVED OVER 200 INTERNATIONAL AND HE HAS OVER 20 BOOKS IN MEDICINE AND THIS NOW LEADS THE REMAINING CLINICAL TRIALS AND APPLICATIONS. SO THIS IS OVER THE YEARS AND DEFINITELY HE'S NEW TO THIS FIELD AND THIS HAS REALLY BEEN MOVING THE BAR AND PUSHING THE BOUNDARIES. WE ARE VERY HONORED TO HAVE HIM TODAY AND WITHOUT ANY FURTHER ADO I WOULD LIKE TO PRESENT THIS TO HIM TO PRESENT THE TITLE MEDICINE APPROACHES TO HEALTHCARE. >> THANK YOU SO MUCH A PLEASURE TO BE WITH YOU TODAY. IT REALLY IS A WONDERFUL PLEASURE TO DO SO AND I'M GOING TO GO AHEAD AND START MY VIDEO AND START THE TALK. IT'S REALLY GREAT TO BE PART OF THIS CONFERENCE WHAT A WONDERFUL ASSEMBLY FOR TALKS THAT WE HAVE PUT TOGETHER. I'M GOING TO GO AHEAD AND SHARE MY SCREEN AND WILL WILL YOU WILL SEE THIS? CAN YOU WILL SEE THE SLIDES? TERRIFIC. THIS IS ACTUALLY A PICTURE OF OUR INSTITUTE AT WAKE FOREST, THE WAKE FOREST INSTITUTE FOR MEDICINE. THIS BUILDING IS BASICALLY FIVE STORIES ABOUT 200,000 SQUARE FEET. EVERYTHING HERE FROM THE CONCEPT TO THE PROOF OF A CONCEPT TO THE ACTUAL BENCHTOP WORK AND THEN WE HAVE OUR OWN PROCESS DEVELOPMENT TEAM AND OUR OWN TEAM THAT BRINGS US TO THE RETAKE REGULATORY CYCLE 58 IN THE MANUFACTURING FACILITY WITHIN THIS BUILDING THAT CREATES PRODUCTS FOR OUR PATIENTS. WE PRODUCE THIS UNDER AN FDA COMPLIANT GMP FACILITY SO WE CAN DELIVER PRODUCTS TO PATIENTS. THE THING WE DON'T DO THIS BUILDING IS TO REACH THE PATIENT. BUT WE DO EVERYTHING FROM THE CONCEPT TO THE CLINICAL STUDIES ALL THE WAY TO THE PRODUCTION OF THE THERAPY. OVER 400 PEOPLE WORK TOGETHER TO BRING THIS TO THAT SIDE. IT IS MY GREAT PLEASURE TO BE ABLE TO SHARE WITH YOU SOME OF THE WORK THAT WE ARE DOING AT THE INSTITUTE. AS YOU KNOW, WE TALK ABOUT REGENERATIVE MEDICINE WITH TALKING ABOUT SIT CELLS AND SCALPELS TOGETHER OR THE SOLE OTHER BUCKETS THAT WE HAVE TECHNOLOGY AND THAT INCLUDES BIO PRINTERS AND BEER BIOREACTORS. THE FIELD HAS BEEN AROUND FOR A LONG TIME. WITH THE FIRST CONCEPT OF THE FIELD IT WAS NOT CALLED REGENERATIVE MEDICINE, BUT THE FIRST CONCEPT WAS AROUND OVER 70 YEARS AGO. SO WHY HAVE WE SEEN THESE CLINICAL ADVANCES IN ALL THESE DECADES. THAT HAS TO DO THREE MAJOR CHALLENGES AND ABILITY TO EXPORT THE CELLS OUTSIDE OF THE BODY AND ADEQUATE MATERIALS ADEQUATE VASCULARITY . MOST OF OUR WORK WE STARTED OVER 30-YEARS AGO NOW WE HAVE TO DO MOST OF IT WITH GROWTH FACTOR BIOLOGY INITIALLY. FIRST THE FIRST GRANT WAS WITH GROWTH FACTOR BIOLOGY. GROW YOUR THREE OH CELLS OUTSIDE OF THE BODY AND THIS GOES TO SELL METHODS AND SELF METRICS INTERACTIONS. THE FACT THIS CANNOT BE EXPANDED IN LARGE QUANTITIES IN THE BODY THIS DUE TO THE FACT THAT SCIENTISTS WERE REALLY TRYING TO GROW THE DIFFERENT CELLS AND THE CELLS THAT WE REALLY NEEDED WITH THE UNDIFFERENTIATED CELLS WE NOW CALL COMMITTED REGENERATIVE CELLS. BY REALLY TARGETING WHAT WE NOW CALL COMMITTED REGENERATIVE CELLS AND EXPANDING THE CELLS OUTSIDE OF THE BODY THEN YOU CAN THEORETICALLY REALLY EXPANDS THE CELLS INTO LARGE QUANTITIES WHICH ARE REALLY THIS NEEDED FOR CLINICAL INFORMATION WHICH IS A CRITICAL EXPANSION STATE AND THAT WAS THE CRITICAL FEATURE TO MAKE SURE THAT WE CAN GET THESE INTO THE PATIENTS. >> INVEST FOR 30-YEARS LATER, MOST HUMAN CELLS CAN BE GROWN AND EXPANDED OUTSIDE OF THE BODY. THERE ARE JUST A FEW BACK 31 YEARS AGO. BUT EVEN NOW, 2021 YOU STILL CANNOT GO TO THE PATIENT TODAY AND GET A BIOPSY FOR THE PANCREAS, NERVES, LIVER AND GET ALL OF THOSE CELL TYPES TO GROW. THERE IS STILL WORK THAT NEEDS TO BE DONE IN TERMS OF CELL BIOLOGY FOR SEVERAL CELL TYPES. THE SECOND CHALLENGE WAS MATERIALS. HOW TO BE DELIVER CELLS INSIDE OF THE BODY? YOU BASICALLY ADHERE TO A VERY SIMPLE PRINCIPLE THAT SCAFFOLDS SHOULD REPLICATE THE BIOMECHANICAL STRUCTURAL PROPERTIES OF THE TISSUE BEING REPLACED. TO PLACE INTO SIMPLE WORDS, IF YOU ARE TRYING TO REPLACE THE BLOOD VESSELS GOING TO REQUIRE A VERY DIFFERENT MATERIAL THAN IF YOU'RE TRYING TO REPLACE A PIECE OF BONE. THEIR CHALLENGE WAS VASCULARITY. AND A FULL CLINIC WAS THE FOUNDER OF INTER GENESIS, WE WERE ACTUALLY HIS FORMATE IN BOSTON. IN 1973, THAT WAS IS A VERY SIMPLE PRINCIPLE. THAT CELLS ALONE COULD NOT SURVIVE IN BONES LARGER THAN 0.3 MILLIMETERS CUBED. THIS IS THE MAXIMUM DISTANCE BY WHICH NUTRITION WOULD CURE BY NATURAL DIFFUSION. SO HOW DOES NATURE SOLVE THIS PROBLEM? YOU NEED TO RESOLVE THIS PROBLEM BY BRANCHING. THE LOWEST FORMS OF LIFE SUCH AS ALGAE WHICH YOU CAN SEE AT THE BOTTOM OF THE OCEAN FLOOR, TO AN INDIVIDUAL TREE THE BRANCHES OF AN ORGAN BRANCHING IS A SOLUTION. THAT IS WHY MANY SCIENTISTS START TO WORK ON SCAFFOLD SYSTEMS THAT THE NAKED EYE COULD LOOK LIKE A PIECE OF YOUR SHIRT OR BLOUSE AND UNDER A SCANNING MICROSCOPE WE HAVE A THREE DIMENSIONAL ORGANIZATIONAL STRUCTURE. MATERIAL SCIENTISTS WORK VERY HARD IN DEVELOPING ALL OF THESE MATERIALS THAT WERE ESSENTIAL REALLY TO BRING THIS TECHNOLOGY FORWARD. YOU CAN SEE HERE THAT THE CELLS CAN WEIGH DOWN ON THESE BRANCHES AND STILL HAVE THE FEROCITY NECESSARY FOR INTER- GENESIS AND MU VASCULARITY. BASICALLY LOOKED AT THIS VERY EARLY ON, WE WERE ABLE TO DETERMINE THAT THE CELLS IN THE SCAFFOLD IN THE CELLS ALONE WERE NOT SUFFICIENT OF COURSE FOR VASCULAR RELATION. THIS WAS DETERMINED BY MANY INDIVIDUALS. BUT THIS WORK THAT WE END UP DOING TO SHOW HOW TO ENGINEER TISSUE THAT WOULD BE WELL VASCULARIZED. YOU CAN SEE HERE ON THE FAR LEFT MUSCLE CELLS, HUMAN MUSCLE CELLS THAT WERE GROWN AND EXPANDED OUTSIDE OF THE BODY AND THEN PLANTED INTO MICE TO AVOID REJECTION. YOU CAN SEE HERE THAT WE'VE IMPLANTED THE MUSCLE CELLS ALONE, NOTHING MUCH WHAT HAPPENED. WE ADDED ENDOTHELIAL SALES A LITTLE BIT OF VASCULARITY, BUT NOT TOO GREAT. BUT AT THE END OF THE MOST POTENT GROWTH FACTOR IT WAS MUCH BETTER BUT THERE WAS NO DOUBT THE BEST VASCULARITY OCCURRED WHEN HE HAD THE MUSCLE CELLS PLUS THE GROWTH FACTORS PLUS THAT BELIAL CELLS CAN REALLY FORM MUSCLE TISSUE THAT WAS RESEMBLING THAT OF A NEGATIVE APPROACH. YOU NEED THAT TO HELP MOVE FORWARD. THIS IS ACTUALLY ONE OF OUR FIRST ATTEMPTS TO DO THIS. THIS IS BY USING ENGINEERING METHODS TO LOOK AT THE URETHRA. THE URETHRA IS A CHANNEL THAT CONNECTS THE BLADDER TO THE OUTSIDE OF THE BODY. WE DID THIS IN RABBIT STUDIES WERE RE- EXCISED A1 CENTIMETER OF THE RABBIT URETHRA AND THEN WE REPLACED IT WITH THE TWO POLARIZED EXTRACELLULAR MATRIX. WE TOOK A PIECE OF BLADDER TISSUE, WE USED DETERGENTS TO WASH THE CELLS AWAY BE STABILIZE THE STRUCTURE AND USE IT TO REPLACE THE ONE SENTIMENT HERE DEFECT IN THE RABBIT. WE DID THIS WITH OR WITHOUT CELLS. LIBBY SHOWED WELL WAS IF YOU DO NOT USE CELLS THEY DID NOT WORK WELL. DUTY TO THE CELLS FOR THIS TO WORK. THE MAXIMA DISTANCE AT WHICH THE CELLS WOULD TRAVERSE THE DEFECT WITHOUT A SCAR FORMATION WAS ABOUT 1 CENTIMETER FROM ANY EDGE. AND SO WE BEEN ENDED UP A FEW YEARS LATER, ABOUT SIX YEARS LATER THIS IS A 1996 WE BASICALLY ENDED UP DOING THIS IN PATIENTS WHERE THE PATIENTS PRESENTED WITH A URETHRAL INJURY. YOU CAN SEE HERE THE INJURED AREA, THIS IS MOSTLY IN THE TOP PORTION OF THE URETHRA AND THEN WE THEN TRIED TO RESPECT PLACES WITH THE SCAFFOLD ALONE WITHOUT THE CELLS. I JUST MENTIONED TO THE METRO DISTANCE FOR MAXIMUM INFUSION THIS DEFECT WAS ABOUT 5 CENTIMETERS OR FOUR AND A HALF CENTIMETER SO HOW DID WE DO THAT? USE THAT BY USING THE SCAFFOLD ALONE AND ONLY REPLACING THE TOP PORTION OF THE CONSTRUCTS. BY DOING SO THE NORMAL TISSUE TO THE CENTRAL PORTION OF THE CONSTRUCT WAS LESS THAN HALF A CENTIMETER FROM ANY EDGE. THEREFORE, THIS WOULD ALLOW THE NEGATIVE CELLS TO WALK ON THAT BRIDGE AND BRIDGE THE GAP. OF COURSE THESE MATERIALS DO YOU SELL YOUR RICE REALLY ABSORB OVER TIME. DURING THAT PERIOD THE CELLS THAT WALKED IN THAT BRIDGE WOULD SENSE THIS MATERIAL WOULD GO AWAY AND THIS WOULD DO THEIR VERY OWN BODY WHICH IS DELAYED ON THEIR OWN MATRIX. SIX-MONTHS LATER YOU ARE LEFT WITH YOUR OWN SELF AND YOUR OWN MATRIX. THIS IS A URETHRA GRANDMA THE SAME PATIENT. HERE YOU SEE THE HISTOLOGY SHOWING THE NORMAL HISTOLOGY OVERALL. AND THEN THIS IS A SCOPIC VIEW SHOWING THE AREA OF THE STRUCTURE BEFORE SURGERY IN THE AREA OF THE REGENERATED URETHRA AFTER SURGERY SHOWING YOU THE NORMAL URETHRA. THAT WORKED FOR SMALL DEFECTS, LESS THAN HALF A CENTIMETER FROM ANY EDGE AND WE WERE ABLE TO DO THAT BUT REPLACING THE TOP TEN OF THE URETHRA. BUT HERE IS A MUCH MORE COMPLICATED PATIENT WITH A MUCH MORE COMPLEX INJURY. YOU CAN SEE A SHATTERED URETHRA WHICH IS MUCH LARGER FROM ANY EDGE. YOU CANNOT CREATE A BRIDGE HERE, YOU HAVE TO CREATE THE ENTIRE STRUCTURE. WHAT DO WE DO HERE? BASICALLY THIS IS OUR ENGINEERING APPROACH WHERE WE TAKE A SMALL SAMPLE OF TISSUE FROM THE PATIENT STRUCTURE AND WE DRAW AND EXPANDED CELL OUTSIDE OF THE BODY. WE CREATE A SIMILAR STRUCTURE WITH THE SCAFFOLD IN BE SURVIVE THE SCAFFOLDS UP THE MUSCLE CELLS ON THE TOP, SEE FIGURE MATERIAL CELLS ON THE INSIDE AND THEN WE THEN PUT IT IN THE BIOREACTOR AND PUT IT BACK INTO THE PATIENTS. AND THAT'S WHAT WE DID IN THIS PATIENT SERIES HERE. ON THE FAR LEFT IS THE PATIENT RANKED ON THE SECOND PANEL YOU CAN SEE THE TWO PLURALIZED SCAFFOLD MADE OUT OF POLLY GLYCOLIC CASON IN POLLOCK LACTIC ACID IN THIS CASE. WE THEN CODED THE OUTSIDE WITH THE MUSCLE CELL WITH THE YOUTH THE REAL CELLS YOU CAN SEE THE INSURED CONSTRUCT TO YOUR RIGHTS WHICH WAS A MATURED REACTOR AND THEN PLANTED BACK INTO THE PATIENTS. YOU CAN SEE HERE THE ACTUAL STRUCTURE BEING IMPLANTED SURGICALLY INTO THE PATIENT AND HERE IS THE PATIENT'S X-RAY SIX YEARS AFTER SURGERY SHOWING THE FORMATION OF THE URETHRA ENGINEERED. WE PUBLISH THAT IN THAT PATIENT SERIES AND YOU CAN SEE HERE THAT PILOT SERIES WITH THESE PATIENTS AND WITH THE INITIAL URETHRAL INJURIES. THE EARLIEST POST OFF APPRAISED FOR THREE MONTHS AFTER SURGERY IN THE LOTUS WINTHROP TO SIX YEARS AFTER SURGERY. WE FOLLOW THESE PATIENTS UP TO SIX YEARS BEFORE HE ACTUALLY PUBLISHED THE PAPER IN THE LANCET. THIS SHOWS THE ACTUAL FLOW OF THESE PATIENTS AND WE NOW HAVE OVER A 20 YEAR FOLLOW-UP USING THE SCAFFOLD ALONE, WITHOUT THE CELLS FOR SMALL DEFECTS REPAIRS UNDER LESS THAN A HALF A CENTIMETER IN OVER A TEN YEAR FOLLOWING THE SCAFFOLD WITH CELLS FOR THE LARGER REPAIRS. WE FELT THAT WE HAVE OTHER STRUCTURES AND BLOOD VESSELS. WE HAVE THE SAME STRATEGY HERE AND INSTEAD OF YOUR OTHER SALES. IF YOU CAN SEE HERE THAT WE CAN BUILD THIS. THIS IS IN THE EARLY 1990s. WE FOUND WHEN WE STARTED TO ENGINEER THESE TUBULAR STRUCTURES IS THAT THEY WOULD COLLAPSE ON THE CULTURE PLATES. THAT IS WHEN WE APPROACH OUR MECHANICAL ENGINEERS AT THE CHILDREN'S HOSPITAL AND MET HARVARD MEDICAL SCHOOL AND WE NEED A DEVICE THAT WILL HELP US TO KEEP THESE THINGS ALIVE AND OPEN. STARTED TO DEVISE THESE BIOREACTORS THAT BASICALLY WOULD ALLOW FLUID TO GO THROUGH THE STRUCTURES AND GET THEM INSIDE OF THE INCUBATOR. THIS IS A CAROTID ARTERY THAT WAS REPLACED USING THESE TECHNIQUES. THIS IS ACTUALLY HIGHLIGHTED IN THIS JOURNAL OF NATURE MEDICINE TO ISSUE THE RESPONSE OF THESE VESSELS AND SO MANY SCIENTISTS HAVE NOW MADE IT SO MANY ADVANCES IN THE AREA OF ENGINEERED BLOOD VESSELS AND WITH CLINICAL TRIALS AND CLINICAL APPLICATIONS OF MANY DIFFERENT LISTED HERE. WE ALSO WORK ON HEART VALVES AND WE HAVE NOT IMPLANTED IN HIS IMPATIENCE YET BUT TO SHOW YOU HOW THESE WORK THE PURPOSE OF THEM IS TO DO AS MUCH AS POSSIBLE TO REPRODUCE A THREE DIMENSIONAL FUNCTIONALITY OF THE STRUCTURE BEFORE THEY GET IMPLANTED INTO PEOPLE. YOU CAN SEE HERE THEY ARE OPENING CLOSING AND SEEING THEIR CELLS SO THEY DON'T GET SLAPPED OFF WHEN THEY ARE CONNECTED TO A HIGH-PRESSURE SITUATION. YOU CAN SEE HERE THAT URETHRAL LAYERING AS YOU USE THESE BIOREACTORS. HERE IS AN INTERESTING ANALYSIS OF PATIENTS AND TECHNOLOGY. WE TOOK THIS ALL THE WAY FROM THE BEDSIDE AS WELL AS THE OTHER ONES. THIS IS USING A VEGETAL ORGAN. NOW COME THE SAME CHALLENGE HERE. WE CANNOT GET THE CELLS TO EXPAND OUTSIDE OF THE BODY. THEY COULD BE HARVESTED AND KEPT ALIVE IN SMALL NUMBERS BUT THE EXPANSION WAS THE KEY. I'M GOING TO USE A SPECIFIC ORGAN TO SEE HOW WE ACTUALLY GO THROUGH THIS PROCESS OF TAKING SOMETHING FROM A BENCH TO THE BEDSIDE. SAME CHALLENGE HERE WE HAPPEN TO THE CELL BIOLOGY THAT TAKES SEVERAL YEARS. TO GET THE RIGHT TO GROWTH CONDITIONS IN THE RIGHT RELEASE CRITERIA SO YOU CAN CREATE THE SAME CELLS TIME AND AGAIN FROM DIFFERENT PATIENTS. ONCE WE WERE ABLE TO AT THE CELLS TO EXPAND OUTSIDE OF THE BODY WE ARE ABLE TO WORK ON THE RELEASE CRITERIA FOR BOTH THE URETHRAL CELLS. WE USE A FAMILY OF OVER 40 DIFFERENT MATERIALS THAT WE MIX AND MATCH THAT WE NEED. WE TALKED ABOUT THIS MATRIX THAT WE HAVE USED AND WE HAVE TALKED ABOUT GLYCOLIC ACID AND FOR THIS PARTICULAR ORGAN WE ACTUALLY USED SIS AS THE EXTRACELLULAR MATRIX. INITIALLY WE USED PG AND PLA AND THEN SWITCH OVER TO SIS FOR THE VEGETAL ORGAN BECAUSE OF ITS ELASTICITY. BUT BASICALLY, ONCE WE KNOW WE CAN GET THE RIGHT RELEASE CRITERIA IN THE RIGHT RELEASE BIOLOGY, IN PARALLEL WE DEVELOPED A BIOMATERIALS STRATEGY AND WE THEN TAKE THOSE HUMAN CELLS AND THEN WE IMPLANT THEM INTO THYMIC MICE TO AVOID REJECTION TO SHOW THE CELLS CAN REALLY READ EVENTUALLY ORGANIZE THEMSELVES. IT WAS WITH US OUR RODENT STUDIES AND WE GO TO THE RABBIT MODEL WE DO WHAT WE DO IN A HUMAN. TAKE A SMALL BIOPSY FROM THE RABBITS, A VEGETAL ORGAN, WE EXPAND THE CELLS OUTSIDE OF THE BODY CAN WE CREATE A THREE-DIMENSIONAL SCAFFOLD. WE THEN DO ' PLACEMENT TO DECEIVE THE CONTRIBUTION IS FROM THE ENGINEERED TISSUE IN THE NATIVE TISSUE AND THEN WE GO TO A FULL TOTAL REPLACEMENT. WE KNOW THE ORGAN THAT WE CAN SEE IS GOING TO BE DEPENDED ENTIRELY ON THE SCAFFOLD ITSELF. WE PUT THIS IN AND THIS IS IN TERMS OF ITS NEGATIVE ENVIRONMENT. THESE ARE THE MORE SOPHISTICATED BIOREACTORS THAT WE USE NOW. THEY HAVE A LOT OF FEEDBACK LOOPS AND WE CAN TARGET THESE AND STOP FOLLOW THESE ALONG AS THE STRUCTURES GET MATURE. THESE ARE THE FULL REPLACEMENT ENGINEERED VEGETAL ORGANS IN THE RABBIT MODEL. TO SHOW THAT IF WE USE THE SCAFFOLDS ALONE, IT'S A MUCH LARGER AND HAVE A CENTIMETER FROM THE EDGE AND IS GOING TO CONSTRICT, CONTRACT, ONLY TO SCAR. IF WE USE THE CELLS THEY FORM A NORMAL ANATOMICAL CONSTRUCT AS YOU SEE HERE IN YOUR GRAPH OVER TIME THIS SHOWS WHAT IS OCCURRING THAT AT ONE MONTH THE CONSTRUCT IS NOT YET COMPLETE. THEY'RE STILL IN BOMB INSIDE THE BODY. YOU CAN STILL SEE THE MUSCLES THAT ARE A THIN LAYER AND YOU CAN SEE THE SCAFFOLD THROUGH A THIN LAYER OF MUSCLE IS STILL PERFORMING. THE MORE MATURE MUSCLE HERE BY THREE MONTHS OF SCAFFOLDS ALMOST AS ALL GONE AND AT THIS TIME THE CELLS THEMSELVES ARE LAYING DOWN THEIR OWN MATRIX. BY SIX-MONTHS WE ARE LEFT WITH THE CELLS IN MATRIX THAT WAS LAID DOWN. IF YOU DON'T USE THE CELLS OF COURSE IT'S GOING TO RESTRICT YOUR STUFF ON THE BOTTOM HALF. THE MUSCLE PHENOTYPE THAT APPROXIMATES THE NORMAL ORGAN BY SIX-MONTHS. WE LOOK AT THE MEXICO GENES AND MARKERS AND PROTEINS APPROXIMATE TO A NORMAL ORGAN BY SIX-MONTHS AND WE LOOK AT THE COLLEGE AND ONE, TWO, THREE AND THE ELASTIN. THIS IS NOW AFTER SIX-MONTHS. YOU ARE SEEING THE MATRIX OF THE CELLS THEMSELVES LAY DOWN AND AGAIN IT APPROXIMATES THE NORMAL ORGAN BY SIX-MONTHS. WHEN YOU LOOK AT THE TENSILE STRAIN AND THE ABILITY OF THESE ORGANS TO COLLAPSE AND AGAIN IT APPROXIMATES THE NUMBER ORGAN BY SIX-MONTHS. WHEN WE LOOK AT THE INNERVATION AND AGAIN IT APPROXIMATES NORMAL ORGAN BY SIX-MONTHS. THIS OCCURS IN CONCERT WITH THE VASCULAR PROCESS THAT OCCURS TOGETHER TO SHOW THAT WE DID IN FACT HAVE INNERVATION AND ITS ENGINEERED TISSUE WE TOOK SAMPLES OF THE VEGETAL TISSUE IN PLACE TO IN VITRO IN A STUDY TO SHOW THAT THIS MIGHT BE HAPPENING FROM THE ENGINEERED TISSUE AND NOT FROM THE SURROUNDING AREAS. THIS CAN SHOW THE INNERVATION OF THESE TISSUES OVER TIME. THIS ACTUALLY SHOWS THE MRI OF THE PATIENT WITH THEM MY ROOKIE SYNDROME. AS YOU KNOW THESE ARE PEDIATRIC PATIENTS WHO WERE DIAGNOSED WITH A CONGENITAL ABSENCE OR ABNORMALITY IN THE ORGAN. THEY HAVE A RUDIMENTARY STRUCTURE YOU CAN SEE WITH THIS MRI ON THE RIGHT AND THE WATER ON THE LEFT AND THE ORGAN WITH A RUDIMENTARY STRUCTURE THAT WE CAN TAKE A BIOPSY FOR THIS STRUCTURE, WE CAN EXPAND THE CELLS OUTSIDE THE BODY AND WE CAN TAKE A THREE-DIMENSIONAL SCAFFOLD WILL BE CREATED, CODES OUTSIDE OF THE MUSCLE CELLS, THE INSIDE WITH THE VEGETAL URETHRAL CELLS. THIS IS A PATIENT ONE YEAR AFTER SURGERY SHOWING YOU THE RECTUM ON THE RIGHTS, FLATTER ON THE LEFT AND THE NEW ENGINEERED VEGETAL ORGAN IN THE SHOWS BASICALLY THE HISTOLOGICAL ANALYSIS OVER TIME. THIS WAS PUBLISHED IN 2014. WE WHEN WE PUBLISH THE WORK WE ALREADY HAVE AN EIGHT YEAR FOLLOW-UP ON HIS PATIENTS. PEOPLE ASKED US WHY DID YOU WAIT EIGHT YEARS? BECAUSE WHEN YOU'RE USING INTESTINE OR SKIN WHICH IS THE CURRENT STANDARD, YOU TEND TO SEE COMPLICATIONS ALL THE WAY OUT TO TWO YEAR FIVE. I WANTED TO MAKE SURE THAT THIS TECHNOLOGY CONTINUED INTO THIS LONG TERM. THESE PATIENTS WERE TESTED OVER AN EIGHT-YEAR PERIOD WITH ENDOSCOPY, BIOPSIES, MOLECULAR TESTING, HISTOLOGICAL TESTING, FUNCTIONAL TESTING AND MRIS SHOW THESE ORGANS CONTINUED TO DO WELL OVER TIME AND THEN SHOWING AGAIN FUNCTIONALLY HOW THESE ORGANS CONTINUE TO DO WELL OVER TIME ALSO. WE ARE CONTINUING THAT WORK WITH CONTINUED STUDIES THAT WE ARE PUSHING THROUGH NOW THE NEXT PHASE. THE FIRST ORGAN WAS PLANTED WAS THE BLADDER. BLOOD VESSELS ARE THE SECOND LEVEL OF HOW THE VASCULARITY IS THE SAME IN ENGINEERED UTERUS AS IT IS THE NORMAL CONTROLLED UTERUS. OF COURSE A SCAFFOLD THAT WAS NOT SEEDED WITH CELLS DID NOT DO WELL. THE SAME THING WITH IMPROVEMENTS IN TERMS OF ENDOMETRIAL THICKNESS AS WELL AS ENDOMETRIAL GLANDS. YOU CAN SEE HERE HOW THAT REALLY IS EQUIVALENT TO THE ENGINEERED UTERUS. THESE ANIMALS WERE THEN ALLOWED TO MOVE FORWARD IN TERMS OF FERTILIZATION AND GESTATION AND WE WERE ABLE TO SHOW THAT THE EMBRYOS DID ATTACHED TO THE ENGINEERED UTERUS WITH DELIVERY OF THE FETAL COUGHS FROM THE BIOENGINEERED WOMB. THIS WAS HIGHLIGHTED IN NATURE ABOUT TECHNOLOGY THIS PAST YEAR. THIS IS STILL A WORK IN PROGRESS THAT IS STILL IN THE PRECLINICAL LEVEL. BY FAR THE MOST COMPLEX ORGANS OR THE SOLID ORGANS, BECAUSE THEY HAVE SO MANY MORE CELLS PER CENTIMETER. SAME CHALLENGE HERE WE BASICALLY HAVE MANY DIFFERENT CELL TYPES BUT TO BEING SOLID, A SOLID STRUCTURE DOES REQUIRE MUCH MORE EFFORT INTO MAKING SURE THAT THIS WORKS. TO SHOW YOU THIS PAPER AND WE WERE REPLACING ENTIRE PENAL STRUCTURES SO WE DID BASIC CELL BIOLOGY STUDIES AND WE DID WE DID THE RODENT STUDIES DANCE WE DO THIS WITH HUMAN CELLS AND THEN WENT TO A RABBIT MODEL WE DID A PROP REPLACEMENT IS A SHOWING OF THE FULL REPLACEMENT. BASICALLY WHAT WE LEARNED HERE EARLY ON IS THAT WE HAD A CHALLENGE IN TERMS OF THE SOLID SCAFFOLDS. AT THE TIME WE DID NOT HAVE THE TECHNOLOGY NECESSARY TO CREATE A SOLID SCAFFOLD AND THAT IS WHEN IT OCCURRED TO US. WHY DON'T WE TAKE A DONOR ORGAN THIS IS A VERY MILD DETERGENT TO WASH THE CELLS AWAY. AND THEN WE RESELL THE RISE THAT SOLID STRUCTURE. THIS WAS IN THE EARLY 1990s ABOUT 1994 WHEN WE CAME UP WITH THIS CONCEPT BECAUSE WE COULD NOT GET THE SOLID STRUCTURE TO WORK. THE STRATEGY IS WE TAKE TISSUE AND EXPAND THE CELLS OUTSIDE OF THE BODY AND WE TAKE A DONOR THAT WE DO SELL YOUR RISE AND TO BE THEN RESELL YOUR EYES THAT WITH THE CELLS AND THEN WE DID THE FULL REPLACEMENT AS YOU SEE HERE. THIS IS UNDER THIS RABBIT MODEL THAT YOU CAN SEE HERE. YOU CAN SEE IN THIS RABBIT'S MODEL, WHICH IS LOOKING AT THE VASCULARITY IN THIS IS ONE OF THE MOST VASCULAR ORGANS IN THE BODY TO SEE YOU THIS IS THE ENGINEERED PENAL TISSUE WITH ITS TWO CORPORAL BODIES AND OF COURSE THE CONTROLS DID NOT DO WELL. WHEN WE LOOK AT THE PRESSURES BY SIX-MONTHS THE PRESSURES WERE SUFFICIENT FOR ERECTION, PENETRATION, COPULATION AND EJACULATION. WE THEN ARE ABLE TO MAKE THESE ANIMALS WITH THEIR FEMALE PARTNERS AND WE WERE ABLE TO TAKE SPERM FROM THE BADGE AND NO FAULT OF THE FEMALE PARTNERS AND THIS LED TO OFFSPRING. WHERE WE ARE WITH THIS TECHNOLOGY, THIS IS WORK THAT WAS FUNDED BY WOUNDED WARRIORS BECAUSE OF THE INCREASED USE OF IMPROVISED EXPLOSIVE DEVICES THIS IS NOW BEEN APPROVED BY THE FDA TO MOVE INTO PATIENTS THAT HAVE PENILE DEFECTS AND THIS IS NOW CURRENTLY ACTIVELY RECRUITING PATIENTS FOR THIS TECHNOLOGY. NOW, WE STARTED WITH THE PENAL TISSUE SO WE QUICKLY MOVED ON TO OTHER SOLID ORGANS LIKE THE LIVER. THIS IS WORK THAT WAS THE HOWARD HUGHES A FELLOW IN BOSTON, SAME STRATEGY HERE. WE TAKE THE LIVER, WE'LL USE A VERY MILD DETERGENT TO WASH CELLS AWAY, TWO-WEEKS LATER YOU HAVE SOMETHING THAT LOOKS LIKE THE LIVER. YOU CAN FOLD IT LIKE A LIVER AND IT FEELS LIKE A LEVER BUT IT HAS NO CELLS. YOU ARE ABLE TO PRESERVE THE VASCULAR TREE AND IS A SHOWING OF THE LIVER HERE. YOU CAN SEE THIS HAS BEEN DAY SELL YOUR RISE AND WE ARE INJECTING CONTRAST INTO THIS DECELERATE HIS LIVER AND YOU CAN SEE HOW THE VASCULAR TREE HAS BEEN PRESERVED. YOU CAN SEE THAT WE CAN NOW INFUSE THAT WITH ENDOTHELIAL CELLS AND PERFUSED THE REST WITH THE REST OF THE LIVER CELLS. THIS WILL END UP WITH A THREE DIMENSIONAL LIVER STRUCTURE THAT'S BASICALLY REPRODUCES THE DEVELOPMENT OF A FETAL LIVER. WHEN YOU LOOK AT IT FETAL LIVER ARE YOU LOOKING AT A SEXTANT DOCTOR LOOKED AND MATURED THE DUCKS AND THAT IS EXACTLY WHAT WE SEE WITH STRUCTURES IN THE DEVELOPMENT OF THE TYPICAL PATTERN THAT IS ABLE TO SECRETE AND METABOLIZE. WE ARE ABLE TO, THIS IS NOT READY FOR PRIME TIME BECAUSE SOLID ORGANS SUCH AS THE LIVER AND KIDNEYS WITH THIS STRATEGY THE CHALLENGE AS OPPOSED TO THE PENAL TISSUE HAS VERY WIDE OPENINGS IS THAT FOR THESE TISSUES THE ENDCAP LAYERS ARE VERY SMALL. WERE NOT THERE YET FOR THESE MORE COMPLEX ORGANS AT THIS POINT. BUT IS STILL IN DEVELOPMENT. THE KIDNEY IS ANOTHER ORGAN THAT WE TARGETED. AGAIN, WE TRY TO USE MANY DIFFERENT STRATEGIES. WE VIEW SCAFFOLD SYSTEMS WE VIEW CELL THERAPY STRATEGIES AND THE KIDNEY IS A VERY COMPLEX ORGAN WE HAVE TO GO THROUGH A LOT OF WORK TO FIGURE OUT MECHANISMS TO GROW AND EXPAND THE CELLS OUTSIDE OF THE BODY. IT TOOK SEVERAL DECADES TO MAKE THAT POSSIBILITY. THIS IS STUDIES WHERE WE ENDED UP CEDING THESE KIDNEY CELLS INTO COLLEGE AND MATRICES THAT WE IN VITRO WITH THESE RENAL UNITS AND WE WERE ABLE TO PANTIES INTO COWS AND STEER SHOWING THAT WE USE CLONE CELLS TO DO THAT FOR THESE INITIAL EXPERIMENTS AND YOU CAN SEE HERE THIS THREE DIMENSIONAL THIS FLUID THAT IS CONSISTENT WITH DILUTED URINE, OF COURSE IF WE USE ENGINEERED KIDNEY THAT WAS NOT HONESTLY OF COURSE IF IT WAS NOT SEATED CELLS WOULD NOT HAPPEN. THESE ARE IMPLANTED INTO COWS AND STEER AND THIS SHOWS THE NEPHRON UNIT WITH THE OWNER LIFE PROXIMAL TUBULES. YOU CAN SEE HERE THE PROXIMAL DISTAL TUBULES HERE AND WHEN WE LOOKED AT THIS LEVEL WITH VITAMIN D3 LEVELS THAT WERE CONSISTENT WITH KIDNEY AND WE LOOKED AT THE MOLECULAR MARKERS AND PROTEINS AND GENES WERE CONSISTENT WITH KIDNEY AND THIS WAS ALSO HIGHLIGHTED IN NATURE BY TECHNOLOGIES WITH SUCCESSFUL TRANSPLANTATION OF CLONE CELLS TO CREATE THEM. WHERE WE ARE WITH THIS TECHNOLOGY IS WE CANNOT GO TO A 70 -YEAR-OLD PATIENT WITH END STAGE AND A BIOPSY FROM THEIR KIDNEY AND THOUGH CELLS CAN THEN BE EXPANDED AND GROW OUTSIDE THE BODY. THOSE CONTAINED IN THE COLLEGE AND CARRY IN THEIR INSERTED BACK. THIS SHOWS YOU HOW YOU ARE ABLE TO RESCUE THESE ANIMALS WITH END-STAGE KIDNEY FAILURE THIS SHOWS YOU HISTOLOGICALLY AND PRETTY CLINICALLY ON THE LEFT THESE TWO UNITS THAT WERE PRESENT ON THEN THEY INSERTED AND THE CELLS FOR THE COLLEGE AND AND YOU CAN SEE ALL THESE NEW NEPHRON UNITS THAT ARE GENERATED AND THIS IS NOW ACTUALLY IN A COMPLETED PHASE TWO CLINICAL TRIAL IN HUMANS ENTERING PHASE THREE CLINICAL TRIALS FOR PATIENTS WHO HAVE END-STAGE KIDNEY DISEASE WHO ARE ON THEIR WAY TO DIALYSIS IN A MANNER TO PREVENT THEM TO GETTING TO DIALYSIS. NOW, ONE OF THE CHALLENGES THAT WE FACE EARLY ON WAS TRYING TO MAKE A LOT OF THESE STRUCTURES BY HAND. TO BE REALIZED EARLY ON THAT TO MAKING IS BY HAND WAS GOING TO BE A CHALLENGE WHEN WE STARTED TO INCREASE THE NUMBER OF PATIENTS THAT NEEDED TO BE TREATED. IT'S OKAY TO MAKE THEM BY HAND FOR A SMALL NUMBER BUT TO SCALE THE TECHNOLOGY UP AND USES COME UP WITH BETTER SOLUTIONS AND THUS WE STARTED THINKING ABOUT 3D PRINTING AND WE STARTED TO LOOK AT, HOW DO WE DO THIS? YOU CAN SEE THIS HERE AND THIS PRINTER WAS MODIFIED WITH THE 3D ELEVATOR AND YOU CAN SEE THIS 3D ELEVATOR HERE SO WE MODIFIED IT EVERY TIME THE PRINTHEAD WOULD GO THROUGH THE STUDIO WOULD ELEVATE IT LOWER ITSELF AND PRINT ONE LAYER AT A TIME. THIS WOULD CREATE THIS THREE DIMENSIONAL HEARTS AND BONE STRUCTURES THAT WE WERE ABLE TO IMPLANT SHOWING FOR THE FIRST TIME THAT YOU COULD REALLY PRINT THE STRUCTURES AND IMPLANTS THEM WITH FUNCTIONALITY. THIS IS THE WORK OF TELLS YOU THAT WAS ONE OF THE FELLOWS IN OUR LAB AND HERE IS THE ACTUAL PRINTER SHOWING YOU THE PRINTING OF THIS TWO CHAMBER HEART AND THIS IS THE ACTUAL TWO CHAMBER HEART THAT WAS PRINTED AT FIRST YOU COULD SEE THE STRUCTURE BEING A RADICALLY BUT THEN IT WOULD START BEATING AND VIEW IT IN UNISON. THIS SHOWS THE FIRST BEATING IN A RADICALLY BEATING IN UNISON AFTER A FEW HOURS. EVEN THOUGH WE WERE ABLE TO SHOW THESE CONSTRUCTS COULD BE PRINTED, THEY DID NOT HAVE THE STRUCTURAL INTEGRITY NECESSARY FOR SURGICAL IMPLANTATION UNLESS WE DID THINGS SUCH AS A BONE. OVER THE NEXT 14 YEARS THEN WE STARTED TO DEVELOP A UNIQUE PRINTING SYSTEM AT THE INSTITUTE THAT WOULD ALLOW FOR TISSUE AND ORGAN CONSTRUCTS TO BE PRINTED WITH THE STRUCTURAL INTEGRITY NECESSARY TO OBTAIN VIABILITY LONG TERM. THIS WAS HIGHLIGHTED IN NATURE BY TECHNOLOGY IN MARCH OF 2016. THERE'S FIVE FEATURES TO THESE PRINTERS. FUTURE NUMBER ONE WE USE SMALL NOZZLES ONTO TO MICRON. TAKE ONE HUMAN HAIR AND SPLIT THAT HUMAN HAIR TEETIMES LODGING TO DELAY THAT IS HOW SMALL WE CAN GET IN TERMS OF THE SOLUTION. THE SECOND FEATURE IS PRECISION. WE CAN PRECISELY DEPOSIT CELLS BUT YET THEY ARE NEEDED. THE THIRD FEATURE IS BASICALLY THE BIO LINKS THAT WE DEVELOPED AND WE DEVELOPED THEM TO GO THROUGH AS A LIQUID BUT ONE THING LED TO THE PLATFORM AND THIS CAN MAINTAIN THEIR INSTRUCTIONAL INTEGRITY THROUGH CROSS-LINKING DIMENSIONS THAT WE USE IT ONCE THEY HIT THAT AND RETAIN THEIR SHAPE. THE FOURTH FEATURE WAS WE CREATED THESE MICRO- CHANNELS THAT WOULD ALLOW FOR NUTRITION TO GO THROUGH TO THE CENTRAL PARTITION OF THE CONSTRUCTS, ALLOWING THEN SOLID STRUCTURES TO BE PRINTED WITH NORMAL MUSCULARITY. THIS WAS IN FACT THE TOPIC OF THE NASA CHALLENGE THAT WAS COMPLETED BY OUR INSTITUTE SHOWING THAT YOU CAN IN FACT PRINT THE STRUCTURES OUTSIDE OF THE BODY AND KEEP THEM ALIVE OUTSIDE THE BODY AS WELL FOR A PROLONGED PERIOD OF TIME OF OVER 30 DAYS WITHOUT HAVING THEM IMPLANTED. BY USING THIS 3D PRINTING STRATEGY. AND THEN THE FIFTH FEATURE WAS TO USE THESE CURRENT AVAILABLE SOFTWARE PROGRAM. IT IS JUST A FAR CRY FROM THE EARLY DAYS WHERE THIS WAS BEING DONE INDEPENDENTLY BY PHYSICIST AND THERE'S NO MAJOR SOPHISTICATED PROGRAM IN EVERY MAJOR HOSPITAL ALLOWS YOU TO THROW IN AN X-RAY TO DETERMINE WHAT THESE ORGANS LOOK LIKE IN PARTICULAR THESE AND PRESIDENT AND EVERY MAJOR HOSPITAL PRETTY MUCH WITH DIFFERENT TYPES OF DIFFERENT COMPANIES BUT WHAT WE DID IS WE DEVELOPED OUR OWN SOFTWARE PROGRAM THAT WE THEN CAN DOWNLOAD THIS DATA FROM THIS IMAGING SYSTEM AND WE CAN THEN CREATE AND PRINT THE STRUCTURE SPECIFICALLY FOR THAT PATIENT. AND THAT IS FEATURE NUMBER FIVE. THIS IS ACTUALLY OUR NEWEST CONCEPT OF PRINTING AT THE PATIENT BEDSIDE AND THIS IS THE ACTUAL SCHEMATIC THIS IS THE ACTUAL PIMP PRINTER ON THE RIGHTS AND THE CONCEPT HERE IS THAT YOU CAN PRINT THIS AT THE PATIENT'S BEDSIDE THIS IS ACTUALLY SHOWING YOU A VIDEO AND THE CONCEPT OF THE PRINTER THAT WE BUILT THAT I SHOWED YOU HERE AND THIS IS SHOWING YOU A PICTORIAL VIDEO OF HOW THIS WOULD WORK. HE SCANNED THE AREA AND AFTERWARDS YOU'RE ABLE TO PRINT THE LAYER WHERE THEY ARE NEEDED WE CAN PRINT THREE-DIMENSIONAL SKINNED WITH MULTIPLE CELL TYPES USING THESE TECHNIQUES. ALSO USING HER BIO PRINTING SYSTEM WE ADVANCE A LOT OF WORK AND THE DIVIDING SHIP AREA WE STARTED THIS WORK SOME TIME AGO, OVER TEN-YEARS NOW. BASICALLY LOOKING AT HOW TO BUILD THESE SYSTEMS AND THE INITIAL WORK WAS FUNDED BY THE REDUCTION AGENCY WITH A 25 MILLION-DOLLAR GRANTS. THE GOAL WAS TO BIO PRINT THESE MINIATURE ORGANS BECAUSE THE MICROCHIP AND TECHNOLOGY AT FOR DRUG SCREENING AND DISCOVERY. SO REALLY IT'S NOT TOO CREATED BY HAND BUT TO BIO PRINT THEM AND THE WAY THAT WE CREATED THESE AS WE STARTED TO WORK ON THESE THE SAME WAY THAT WE ENGINEERED ORGANS. YOU WANT ALL CELL TYPES THAT THE PERSON IN THE ORGAN IN THE SAME PROPORTIONS WITH THE EXTRA CELL MATRIX. SO TO CREATE A LIVER EQUIPMENT FOR EXAMPLE, THE LIVER HAS FIVE MAJOR CELL TYPES, THEY USE AS A PROPORTION AND WE USE THEN USE THE EXTRACELLULAR MATRIX TO THEN CREATE A POWDER WITH IT AND THEN USE AN EXTRACT. WE MIX THEM TOGETHER. PERCEIVING STIFFNESS AS IMPORTANT AS WE CREATE THESE MINIATURE STRUCTURES ABOUT THE SIDE OF A PINHEAD EACH. THIS IS VERY IMPORTANT THAT WE CAN PRODUCE THIS AND BY HAVING THIS SYSTEM WE ARE ABLE TO HAVE A LIVER STRUCTURE WITH HIGH FIDELITY WITH THE HUMAN EQUIVALENT, INCLUDING THE PRESENCE OF MICRO ARE BIO NUCLEAR NUCLEI AS YOU SEE HERE. YOU KEEP THE STRUCTURES ALIVE LONG TERM. THIS IS WITH THE PRODUCTION OF YOUR REAL ANNA BE HUMAN AND YOU CAN SEE HERE A SAME DRUG DOSE RESPONSE THAT WAS EQUIVALENT TO THE HUMANS. THIS SYSTEM CAN BE USED FOR HEALTH IN THE DRUG DEVELOPMENT PIPELINE. USING ALL KINDS OF ORGAN NOISE INCLUDING ORGAN OR VARIANTS THAT I'VE SHOWN YOU. YOU'VE ALSO USED THESE TWO IMPLANTS AND TO SHOW FUNCTIONALITY. WE'VE TAKEN THESE BIOENGINEERED OVARIAN CONSTRUCTS THAT WERE MICRO ENCAPSULATED. THEY WERE IMPLANTED IN RATS WITH THE OVARIES WERE REMOVED, SHOWING A RETURN OF HORMONAL FUNCTION. YOU CAN SEE HERE FOR EXAMPLE PLASMA FSH AND LH AND YOU CAN SEE HERE THE NORMAL CONTROLS AS WELL AS THE ANIMALS THAT WHEN IMPLANTED WITH THIS BIOENGINEERED OVARIAN CONSTRUCT AND SEE THE NORMAL FSH AND LH THE APPROXIMATION OF ESTRADIOL AND PROGESTERONE USED IN THE SAME STRATEGY IN THESE ANIMALS THAT HAD THEIR OVARIES REMOVED. THIS WAS DONE IN THE RAT MODEL BUT THE QUESTION IS, WHAT CAN WE DO IN HUMAN? CAN WE ACTUALLY START TO GET STEM CELLS FROM HUMANS TO TRY TO GET STEM CELLS FROM POSTMENOPAUSAL OR PATIENTS WHO HAD INJURY OR TRAUMA? CAN WE ACTUALLY GET STEM CELLS TO ACHIEVE THE PRODUCTION OF THE FERTILIZED. THIS IS A HUMAN OVARY FROM A DONOR IN THE 20s AND WHAT WE NEED IS NO FOLLICLES NO CELLS TO CREATE THIS AND IF WE DO INSTEAD WE DERIVE STEM CELLS, A STEM CELL POPULATION FROM THIS HUMAN OVARY THAT IS POSTMENOPAUSAL. WE THREW THAT WE BASICALLY GO THROUGH A FOUR STAGE APPROACH AND IN THIS FOUR STAGE APPROACH WE BASICALLY CREATE IN VITRO WHERE WE ISOLATE THE STEM CELLS AND THIS IS A SOMATIC STEM CELLS FROM THE OVARY. WE ARE TRYING TO BASICALLY GET BECAUSE CELLS. [INAUDIBLE] ARE GETTING STEM CELLS AND WE ARE PRINTING THEM TO GRANULES THE CELLS AND WE CAN EXPAND THE SUM CELLS AND PUT THEM INTO FUNCTIONAL OVARIAN CELLS AND THEN WE CAN BIOENGINEERED THESE FOLLICLES USING OVARIAN CELLS AND THEN WE CAN INDUCE THIS IN VITRO OR GENESIS USING CYCLICAL LEVELS OF HORMONES WE'VE GONE THROUGH ALL OF THE STEPS AT THIS POINT. THIS IS USING GERMLINE STEM CELLS AND THE SOMATIC STEM CELLS FROM THE OVARY. THIS ACTUALLY SHOWS YOU THE SITES FROM THE BIOENGINEERED FOLLICLE THAT IS COMPARED HERE TO THE SITES FROM THE NATURAL AND BIOENGINEERED AND THIS IS NOT BIOENGINEERED AND WE ARE NOW AT THE STAGE WHERE WE HAVE THIS APPROVAL TO MOVE FORWARD WITH THIS. AT THIS POINT WE ARE JUST DOING MOCK TESTING AND WE WANT TO MAKE SURE THAT THIS IS ABLE TO BE FORMED. ALL OF THESE STRUCTURES CAN BE FORMED JUST FROM SEPARATE SINGLE STEM CELL POPULATIONS THAT I JUST MENTIONED. WE CAN USE THIS TO CREATE THIS SET GOES THROUGH MITOSIS. THE FIRST STATE OF MEIOSIS ON THE WAY TO THE SECOND STAGE. THIS IS TO CREATE THESE SITES FOR THIS PROCEDURE THAT THE NEXT STEP FOR US. IN TERMS OF CELL THERAPY, WE HAVE LOOKED EXTENSIVELY AT THE CELL THERAPY AS WELL. THE CELLS THAT WE ARE USING IN THE PATIENT'S OWN CELLS TO AVOID REJECTION, IN THIS PARTICULAR CASE FOR EXAMPLE WE TAKE A BIOPSY FROM THE SKELETAL MUSCLE, WE EXPAND THE CELLS OUTSIDE OF THE BODY AND WE CAN ALSO USE THEM AS THE POINT OF CARE AND INJECT THEM BACKED INTO THE DAMAGED REGION AND THE PATIENT. THE SO FOR PATIENTS WITH YEAR-TO-YEAR INCONTINENCE AND WE COMPLETED A TRIAL WITH URINARY INCONTINENCE IN FEMALE PATIENTS, WITH PELVIC HEALTH DISEASE AND WERE CURRENTLY USING THIS TECHNOLOGY NOW WITH PATIENTS WITH ROTATOR CUFF INJURY WHERE THEY HAVE SHOULDER INJURIES IN A CLINICAL TRIAL RIGHT NOW AND WAKE FOREST WHERE WE ARE INJECTING THE CELLS AT THE TIME OF THE REPAIR TO ENHANCE MUSCLE REGENERATION. BASICALLY THE CELLS HAVE HAD EXTENSIVE WORK WITH SKELETAL MUSCLE CELLS THAT ACTUALLY DO VASCULAR. [INAUDIBLE] FUNCTIONALITY. IN TERMS OF STEM CELLS, WE'VE ALSO LOOKED AT STEM CELLS AND AMNIOTIC FLUID FROM THE PLACENTA THAT WE POSTULATED ABOUT 20-YEARS AGO NOW. WE POSTULATED THAT PERHAPS THERE WOULD BE A STEM CELL POPULATION IN THE AMNIOTIC FLUID IN THE PLACENTA. AS YOU KNOW, HUMAN RUNNING STEM CELLS GET ISOLATED BY COMBINING THE SPERM AND THE EGG AND ISOLATING THOSE CELLS 20 DAYS AFTER THE COMBINATION OF THE SPERM INTO THE EGG. OUR HYPOTHESIS WAS, WHAT IF WE JUST WAIT AN EXTRA EIGHT-WEEKS? AND AN EXTRA SEVEN-WEEKS AND THEN TRY TO SEE IF WE CAN GET THE SAME CELL POPULATION FOR THE AMNIOTIC FOOD IN THE PLACENTA. IT WAS A FISHING EXPEDITION BUT IT TURNED OUT TO BE SUCCESSFUL TO OUR SURPRISE WE WERE ABLE TO ISOLATE HIGHLY MULTI- POTENT STAMPS. BUT HIGHLY MULTIPOTENT STEM CELLS FROM BOTH THE AMNIOTIC FLUID IN THE PLACENTA THAT WE WERE ABLE TO BE DIFFERENTIATED INTO ALL THREE GERM LAYERS, ECTODERM, MESODERM AND ENDODERM. THESE ARE COMING FROM BOTH THE AMNIOTIC FLUID AND SAMPLES. THIS IS SHOWING YOU A NORTHERN BLOCK TO SHOW YOU THAT ONE CELL TO GIVE RISE TO ALL THREE GERM LAYERS OF THE CELLS WERE IN FACT MOSTLY POULTON'S AND GOING INTO ALL THREE GERM LAYERS. THEY HAD MARKERS CONSISTENT WITH HUMAN EMBRYONIC STEM CELLS LIKE AQUAPHOR AND SOCKS TOO. BUT THEY ALSO HAD MARKERS CONSISTENT WITH ADULT STEM CELLS WHICH WAS A SURPRISE. WE DID THIS AND THE CELLS KEPT GROWING AND GROWING AND PRESERVED THIS TO THE IPS STEM CELLS BUT THIS IS THE ONE MAJOR DIFFERENCE BETWEEN THE CELLS AND HUMAN EMBRYONIC CELLS. AS YOU KNOW BY DEFINITION HUMAN EMBRYONIC IN THOSE CELLS BUT THE DIFFERENCE HERE IS THAT THE CELLS ARE SLIGHTLY MORE MATURE, YOU'RE GETTING THEM SEVERAL WEEKS LATER IN GESTATION THEREFORE THOSE CELLS DO NOT FORMATS. THAT IS THE MAJOR DIFFERENCE WITH THE CELLS. THESE CELLS CAN BE USED ALSO LIKE ADULT STEM CELLS. THEY CAN BE USED INTO A MODULAR TORY PATTERN OF THERAPY BECAUSE THE CELLS ARE CLONAL YOU CAN GET THE SAME RESPONSE EVERY TIME OR YOU CAN USE THEM FROM MEDICINE APPLICATIONS OR YOU CAN DIFFERENCE THE CELLS FOR THERAPY. THIS TWO WAYS TO USE THE CELLS THIS CAN BE OBTAINED FROM THIS CART THE AT THE TIME OF BIRTH EQUALLY TO THE BABY IS BURNED BORN. THEY DO NOT FORM TARA THOMAS AND LIKE HUMAN AND EMBRYONIC STEM CELLS, UNLIKE ADULT STEM CELLS THAT CAN BE RAPIDLY EXPANDED INTO LARGE QUANTITIES AND GO TO ALL THREE GERM LAYERS. THEY CAN BE STORED FOR SELF FIERCE OR THEY CAN BE STORED IN LARGE QUANTITIES. NO CELLS PERFECT AND NEITHER IS THIS ONE. THE CHALLENGE OF THE CELL IS THAT YOU HAVE TO WORK HARDER TO DIFFERENTIATE. FOR EXAMPLE, YOU CAN TURN THE CELLS INTO LIVER WITHIN TEN-DAYS. WITH THE CELLS IT TAKES ABOUT 20 DAYS TO GET TO THE LIVER AND IT'S ALSO THE SAME PROPERTY THIS ALLOWING YOU TO FORM THIS. THE ADVANTAGE IS THEY DON'T FORM TUMORS. THIS WORK WAS ALSO HIGHLIGHTED IN TECHNOLOGY STEM CELLS FROM THE WOMB. THIS IS NOW MOVING FORWARD TO THE CLINICAL TRIALS AS WELL. TO SUMMARIZE WHAT I'M TRYING TO DO IS GIVE YOU AN OVERVIEW OF USING DIFFERENT STRATEGIES USING MATERIALS ALONE WITH SMALL DEFECTS FROM THE EDGE USING THE CELLS TOGETHER FOR OUR ENGINEERED CONSTRUCTS WITH TISSUES AND ORGANS USING CELLS ALON ALONE, ALTHOUGH WE HAVE USED SMALL MOLECULES AS WELL WHICH I DID NOT PRESENT TODAY WE USED DIFFERENT STRATEGIES SUCH AS 3D PRINTING OR USING SOLID CARD ORGANS WHICH WE DECELERATE THE RESELLER RISE AND WE ALSO TALKED ABOUT THE COMPLEXITY, LEVEL ONE BEING THE LEAST COMPLEX LIKE STRUCTURES AND LEVEL TWO BEING ANOTHER LEVEL OF COMPLEXITY WITH A TUBULAR ORGANS SUCH AS BLOOD VESSELS, LEVEL THREE BEING HOLLOW OR TUBULAR ORGANS SUCH AS VEGETAL ORGANS AND EVENT LEVEL FORCE BEING SOLID ORGANS SUCH AS THE KIDNEY AND NOW WE HAVE BEEN ABLE TO IMPACT ALL FOUR AREAS CLINICALLY USING THESE DIFFERENT STRATEGIES., THE MAJOR MESSAGE HERE THIS IS A REQUIRED MULTIDISCIPLINARY TEAM. MICROBIOLOGIST, CELL BIOLOGIST, BIOENGINEERS, THIS ALL IS EVERYBODY WORKING TOGETHER TO BRING THESE TECHNOLOGIES TO THE BEDSIDE IT REALLY TAKES A VILLAGE AND IT TAKES DECADES OF WORK. AS YOU CAN IMAGINE EVERYTHING I SHOWED YOU TODAY WORKED BUT THAT'S BECAUSE WE WENT THROUGH TIME AGAIN THE ERROR AND THE WORK THAT I JUST SHOWED YOU TODAY SO THAT WORKED 18 YEARS AGO TO GET TO WHERE WE ARE TODAY. THESE ARE VERY, VERY COMPLEX PROGRAMS THAT NEED LONG-TERM FOLLOW-UP WITH THE TEAM AND WERE VERY FORTUNATE TO HAVE AN AMAZING TEAM OF FACULTY WORKING TOGETHER TO BRING THESE TECHNOLOGIES TO THE BEDSIDE. THIS MULTIDISCIPLINARY APPROACH WE TRY TO PRODUCE BY THE ARMED FORCES AND THIS IS A MAJOR EFFORT TO BRING UNIVERSITIES TO WOUNDED WARRIORS AND HAVE AN AMAZING TEAM OF INVESTIGATORS. BILL WAGNER, JAMES TWO, JEFF GARDNER, RICHARD KLOCK, AND SEAN JACKSON ET CETERA THESE ARE THE LEADERS NATIONWIDE THAT HELP US RUN THIS PROGRAM HERE OVER THE INSTITUTE WAKE FOREST AND TO BRING A TEAM TECHNOLOGIES TO PATIENTS THROUGH THIS EFFORT. AT THIS TIME I WOULD LIKE TO THINK OUR FUNDING SOURCES WHICH MAKE THIS WORK POSSIBLE AND FINALLY TO THANK ALL OF YOU WITH YOUR ATTENTION TODAY AND AGAIN FOR THE INVITATION AS WELL AS YOUR TEAMMATE BAYER AND IT'S REALLY A PLEASURE TO BE WITH ALL OF YOU TODAY. THANK YOU IT'S NICE TO BE ELEVATED THANK YOU. >> THANK YOU. UNFORTUNATELY, THE THIS IS DEFINITELY AN ENLIGHTENING PRESENTATION IS VERY INSPIRING AND CONGRATULATIONS TO YOU AND TO YOUR TEAM. IT SHOWS THAT IT TAKES PERSEVERANCE, A LOT OF WORK AND IF YOU ARE SET TO MAKE CHANGES YOU HAVE THE MINDSET AND YOU'RE READY TO GO THROUGH THIS TRIAL AND ERROR AND YOU SURROUND YOURSELF BY YOU CAN TAKE EXTRA WITH CAPITALISTS AND WE ARE RECEIVING QUESTIONS THROUGH THE VIDEOCAST LINK AND I WOULD LIKE TO GO TO THE PEOPLE WATCHING ON THE BROADCAST TO SEND US THOSE QUESTIONS AND LET'S TALK WITH THE FIRST ONE IS QUESTION RELATED THE MOST MUSCULARITY. CAN YOU DISCUSS WITH US SOME EXAMPLES OF MATERIALS THAT ARE EXCLUSIVE TO MUSCULARITY AND WHETHER OTHER IDEAS SHOULD NOT BE CONSIDERED BECAUSE ALSO, THIS IS A LONG QUESTION AND THIS PROCESS IS THE OTHER WAY. >> SOUNDS GREAT THANK YOU. I THINK THAT QUESTION CAME SO MARTHA THANK YOU SO MUCH FOR THAT QUESTION APPRECIATE IT WE DO MISS SEEING YOU IN PERSON. WE MISS SEEING. >> I AGREE. IT'S GOOD TO SEE YOU. >> LIKEWISE MARTHA THINK IS SO MUCH. THAT IS A VERY GOOD QUESTION BECAUSE IN REALITY THE CELLS OF COURSE ARE YOUR NUMBER ONE SOURCE OF YOUR CELLS RELEASE CRITERIA FOR YOUR CELLS ARE GOING TO MAKE THAT TISSUE AND LAID ON THAT MATRIX. BUT THE BIOMATERIALS ARE JUST AS IMPORTANT. THE BIOMATERIALS ARE SO INCREDIBLY IMPORTANT BECAUSE IT'S A FINE BALANCE I MENTIONED WE HAVE A FAMILY OF OLDER 40 THAT WE USE TO MIX AND MATCH AND WHAT WE ARE TRYING TO ACHIEVE IS FOR TRYING TO ACHIEVE A BIOMATERIAL THAT WILL RECORD LOCATES THE OF THE MATERIAL BEING REPLACED BUT EVEN MORE IMPORTANT IS YOU HAVE TO MAKE SURE THAT THE BIOMATERIAL DEGRADES DOES NOT DEGRADE TOO SOON OR TOO LATE. THAT PERIOD IS THE MOST IMPORTANT THING IS WHAT WE DO AND I WILL EXPLAIN WHY. THE MATERIAL DEGRADES TO QUICKLY YOU'RE NOT GETTING A CHANCE FOR CELLS TO FORM A NEW TISSUE THAT IS NECESSARY AND FOR THE CELLS TO DIFFERENTIATE TO THE POINT CAN LAY DOWN THEIR OWN MATRIX. THE FILE MATERIAL IF IT COULD GET CREATED TOO QUICKLY IT CAN COLLAPSE AND FAIL. ON THE OTHER HAND IF YOU HAVE THE MATERIAL THAT DEGRADES TOO SLOWLY THEN YOU END UP WITH SCAR FORMATION BECAUSE WHAT HAPPENS IS AS WHAT HAPPENS IS YOU HAVE TO DEAL WITH NOT JUST INFLAMMATORY RESPONSE IS OF THESE MATERIALS, BECAUSE YOU'RE STAYING BEHIND BUT YOU'RE ALSO DEALING WITH IT MATRIX OF BEING DEPOSITED BY THE CELLS THEMSELVES COMING IN CONFLICT WITH THE MATERIAL ITSELF SO THAT LEADS TO SCAR FORMATION. SO WITH THAT LUCKILY, THAT PERIOD IS ACTUALLY FIRMLY BROUGHT AND THE TIME THAT YOU COULD USE THESE MATERIALS, YOU DON'T DO THEM TO DEGRADE TOO QUICKLY OR TOO LATE THAT'S ONE OF THE MAJOR THINGS HERE IN THE OTHER THING IS WE'VE USED THESE MATERIALS TO CREATE FLAT TUBULAR AND SOLID STRUCTURES. SO FOR EXAMPLE FOR NASA VASCULAR CHALLENGE, USE OR BUY A PRINTER TO DO THAT AND WE USE THAT FOR THE LIVER AND AGAIN, WE TRY TO USE MATERIALS THAT CAN BE COMPATIBLE LONG-TERM AS WELL AS MATERIALS THAT HAVE THE LEAST INFLAMMATORY RESPONSE FOR SOLID ORGANS AND MISSIONS ARE MENTIONED PENILE TISSUE WHICH IS AN ORGANS OF BEEN TAKEN TO THE CLINIC AND WERE TO TAKE THAT THEY ARE. THE REASON WHY THAT SCAFFOLD WORKS FOR THAT TISSUE IS BECAUSE THE CORPORAL KATHERINE NOSA, SO YOU KNOW WHAT WE'RE DOING FOR THAT, WERE TAKING THE ORGAN IS GIVING AT THIS IDEA BACK IN 1994 IF USING A SOLID STRUCTURE THAT WE DO SELL YOUR EYES. IT WORKS NICELY BECAUSE THE PENILE STRUCTURE WHICH IS A SOLID STRUCTURE IS BASICALLY A SPONGY TISSUE IS JUST A BIG SPONGE RIGHT. THE ABILITY OF US TO USE THAT TO RESELL YOUR EYES IS VERY EASY TO DO IN THE CELLS ARE GOING TO THE ENDOTHELIAL CELLS IN THE SMOOTH MUSCLE CELLS ARE WORKING IN UNISON TO GATHER TO CREATE THE ERECTILE TISSUE. THAT IS MUCH DIFFERENT THAN THE LIVER THAT I SHOWED WITH THE DECELERATES LIVER. THEY DECELERATES KIDNEY THAT WE'VE ALSO TACKLED OR THE DECELERATES TO HEART AND THE REASON FOR THAT IS BECAUSE WHEN YOU'RE TALKING ABOUT THE DECELERATES LIVER OR KIDNEY YOU'RE RELYING ON THE VASCULAR NETWORK THAT INCLUDES A MAJOR COMPONENT IS CAPILLARIES. AS OPPOSED TO THE CARBONATE SPONGY TISSUE OF THE PHALLIC STRUCTURE WHICH IS A SOLID ORGAN. BUT TWO DIFFERENT TYPES OF ORGANS. IN FACT, THAT IS WHY THEY ARE IN THE NASA VASCULAR CHALLENGE WE WENT THROUGH THOSE DIFFERENT APPROACHES BECAUSE ONE OF THEM WAS THIS APPROACH OF THE SPONGELIKE TISSUE AND ONE WAS THE APPROACH OF THE VASCULAR TUBULAR STRUCTURE. SO FOR SOLID ORGAN, RIGHTS AND NOW THE LIMITATION FOR THE SELLER'S MATRICES IS THAT WHEN YOU ARE DECELERATES IN THE SOLID MATRIX THERE IS AND YOU ARE ACTUALLY BRINGING THOSE CELL ELEMENTS AND THEY BASICALLY CAN INCLUDE ROKU THE END CAPILLARIES AND THAT IS THE CHALLENGE WITH THOSE. SO IT REALLY THAT IS WHY FOR THOSE SOLID STRUCTURES WE HAVE NOW JUMPED TO BIO PRINTING WHERE WE CAN ACTUALLY CREATES THOSE FINE TUBULAR STRUCTURES THAT ALLOW US TO CONTROL THE TUBULAR STRUCTURES THAT ARE NOT GOING TO COLLAPSE AND THEY'RE GOING TO ALLOW FOR THAT NUTRITION TO GO THROUGH AND THE PRINTER GIVES US THAT PRECISION THAT ALLOWS US TO DO THAT. AGAIN, THE MATERIAL USED MATERIALS AND WERE NOT CHANGING THE MATERIALS WERE JUST CHANGING THE STRATEGY THAT WE USE FOR THE ENGINEERING. THAT IS A LONG ANSWER BUT IT'S A GREAT QUESTION. >> SO BIO PRINTING IS CRITICAL TO ADVANCES IN THIS FIELD AND IN TERMS OF HAVING CELLS AND MATERIALS TOGETHER TO ADVANCE WHAT IS GOING ON. I DON'T WANT TO CLOG UP TIME. I'M SURE THERE'S OTHER QUESTIONS SO THERE'S OTHER QUESTIONS PLEASE GO AHEAD. >> WE WANT THIS TO BE A DIALOGUE SO BRING IT ON IF YOU HAVE QUESTIONS. THANK YOU THIS IS IMPORTANT THAT WE TALK ABOUT THOSE AND WE APPRECIATE THAT. SO SOMETHING THAT IS SOMETHING I WANT TO TOUCH UPON IS TO TRY TO IDENTIFY WHAT ARE THE ROADBLOCKS? YOU HAVE BEEN VERY SUCCESSFUL WITH THIS VERY LONG TERM PLAN OF MOVING THINGS AND AN IDEA TO TREATING PATIENTS, BUT DO YOU HAVE ANY PERSPECTIVE OF RIGHT ARE THESE ROLES THAT YOU CAN SEE AND MAKING IT MORE EFFICIENT TO BEING A SOLUTION? WILL A PRINTING SOLUTION IN BRINGING THIS TO THE PATIENTS FASTER THIS WAY WE CAN LEARN FROM THE FDA IN THE MANUFACTURING AT THE SAME TIME AND THE PATHWAY IS BECOMING MORE ACCESSIBLE AND THE CHALLENGES ARE IN PART RELATED TO THIS MARKER AND CAN HELP THE VALIDATION SO YOU CAN FULFILL THIS AND THEN LATER SAY IN ORDER TO ALLOW YOU TO MOVE FORWARD. BUT YOU ARE ONE OF THE PIONEERS IN THIS FIELD SO PLEASE TELL US WHAT IS THE SECRET AND WHAT WE NEED TO THINK MOVING FORWARD TO REALLY MAKE IT EASIER FOR HAVING SOLUTIONS MOVING FORWARD. >> THANK YOU ANTONELLO. IT'S INTERESTING RIGHT, I MENTIONED IT JUST TAKES SO MUCH TIME REALLY TAKES DECADES TO BRING THESE PRODUCTS ALL THE WAY THROUGH IT REALLY DOES AND THE REASON FOR THAT IS ENGINEERING AND IT TISSUE ORGANISM IS SUCH A COMPLEX THING. YOU REALLY ARE TRYING TO REPRODUCE THE CELL BIOLOGY, THEOLOGY, FROM ECOLOGY AND THE OTHER PROPERTIES. IT'S A PROCESS THAT YOU HAVE TO DO OVER AND OVER AND OVER AGAIN. YOU COULD DO THIS UNTIL YOU GET EVERYTHING RIGHT. IT MAY BE ALL OF A SUDDEN THAT YOU GET THE STRUCTURE TO HAVE YOUR TISSUE-ENGINEERED CONSTRUCT TO HAVE STRUCTURAL INTEGRITY NECESSARY BUT GUESS WHAT. PHYSIOLOGICALLY IT'S NOT DOING AS WELL BECAUSE YOU CAN CHANGE A PART OF THE FORMULA. GETTING THE FORMULA RIGHT IS THE MOST IMPORTANT THING AND WE ARE DOING THIS WITH BIOLOGICAL SYSTEMS. THIS ARRANGE ALSO OF RESPONSE IS FROM PATIENT TO PATIENT THAT IS THE OTHER CHALLENGE THAT YOU'RE DEALING WITH PATIENT AND PATIENTS IN YOUR CRITERIA HAS TO BE WELL-DEFINED, MANY PEOPLE, THIS IS INTERESTING BECAUSE MANY PEOPLE SAY OH TRANSLATIONAL RESEARCH GUESS WHAT, IF YOU'RE DOING TRANSLATIONAL RESEARCH OR BASIC SCIENCE BETTER BE AS SOLID AS IT CAN BE. IT HAS TO BE MORE SOLID THAN ANYTHING ELSE BECAUSE IF YOU ARE BUILDING THAT FOUNDATION ON YOUR BASIC SCIENCE YOU'RE NOT GOING TO BE ABLE TO GET THERE IN LESS YOUR BASIC SCIENCES SOLID. THEREFORE THOSE KEY ELEMENTS, THOSE ELEMENTS OF YOUR CELL BIOLOGY AND ALL OF THOSE ELEMENTS HAVE TO BE SOLID REPRODUCIBLE TIME AND TIME AGAIN AND AROUND EACH ONE OF THEM YOU HAVE TO BUILD RELEASE CRITERIA THAT WILL BE BROUGHT IN US ALL OF YOUR PATIENTS CAN BENEFIT I THINK THE COMPLEXITY OF THE FIELD IS INHERENT IN ITS GOAL, WHICH IS REGENERATIVE MEDICINE FOR PATIENTS AND IT'S NOT AS EASY AS PEOPLE THINK. THAT'S ONE OF THE THINGS THAT I LAUGH ABOUT WHAT PEOPLE SAY, WE ARE GOING TO USE STEM CELLS AND SECURE EVERYTHING. THE STEM CELL POPULATION IS GOOD FOR THIS IN THE SPIRIT AND IT'S JUST NOT THAT EASY. IF IT WERE THAT EASY WE WOULD BE THERE ALREADY I JUST THINK IT TAKES A LOT OF HARD WORK AND I THINK WE HAVE TO BE VERY CAREFUL ABOUT THE HYPE BEHIND THE FIELD BECAUSE SOMEBODY PUBLISHES SOMETHING IN A MOUSE MODEL AND ALL OF A SUDDEN THAT'S THE CURE FOR HUMANS BUT AS YOU KNOW EVERYTHING CAN BE CURED IN A MOUSE SO I THINK THE CHALLENGES THAT WE HAVE IN THE FIELD REALLY HAVE TO DO WITH GETTING BACK TO BASICS TO MAKE SURE THAT YOUR SCIENCE IS SOLID AS YOU MOVE FORWARD WITH THESE TECHNOLOGIES THE OTHER PIECE IS THE MANUFACTURING THAT YOU MENTIONED BECAUSE RIGHT NOW THE CHALLENGE FOR THE FIELD IS REPRODUCIBILITY AND COST REDUCTION AND RELIABILITY. THAT IS WHERE THE MANUFACTURING BECOMES CRITICALLY IMPORTANT FOR THE SCALE AND BRINGING COSTS DOWN AND WERE NOT THERE YET WE NEED TO BRING THESE COSTS FURTHER DOWN. WE HAVE TO PUT BALANCE OUT TO BECAUSE FOR SOMETHING LIKE THAT KIDNEY WHERE WE ARE RIGHT NOW ENTERING PHASE THREE CLINICAL TRIALS FOR THE KIDNEY. KEEPING A PATIENT ON DIALYSIS IS ABOUT ONE QUARTER OF A MILLION DOLLARS PER YEAR. ARE YOU WILLING TO SPEND $100,000 IN THE CAMP KIDNEY CONTRACT? YES YOU ARE BUT I THOUGHT THE SAME FOR SKIN, WE CAN GET THAT FROM A CADAVER DONOR OR DONOR CADAVER. I THINK THAT THERE'S MANY CHALLENGES OF BASIC SCIENCE AND MANUFACTURING AND ECONOMICS BEHIND ARE THE THREE MAJOR CHALLENGES. REGULATORY CHALLENGES ARE ALWAYS GOING TO BE THERE BECAUSE YOU HAVE THE .-DOT YOUR EYES ACROSS HER TEETH BUT THAT'S WHAT YOU'RE DOING IT'S THE RIGHT THING BY THE FDA AND THE PATIENT BY OBSERVING THOSE PRINCIPLES. I DON'T CONSIDER THAT A CHALLENGE. [INAUDIBLE] >> I YOU HAVE MITIGATED ALL THIS SO IT'S IMPORTANT THIS IS A PARTNERSHIP GETTING THE RIGHT PRODUCT WHEN THE TIME IS RIGHT OR THE RIGHT PATIENTS. >> EXACTLY. THE REGULARS JUST WANT YOU TO DO THE RIGHT THING FOR THE PATIENT AND SO DO WE. I AM CURIOUS ABOUT YOUR SPIN ON ORGANIZE AND HOW NOW WE HAVE TOOLS THAT CAN BE VERY INFORMATIVE ON TOXICITY AND WERE NOT AVAILABLE FOR YOU CAN USE IN THE HUMAN AND THE INFORMATION WE HAVE IS ALSO FOR WHAT ADULTS USE WERE NOT USING STEM CELLS AND IS THERE ANYWAY THAT THAT YOU CAN TAILOR THAT TO LEARN MORE ABOUT THE SAFETY AND PATIENTS THAT NOW HAVE DIFFERENT CHALLENGES THAT PEDIATRIC PATIENTS GO FROM ZERO TO PREADOLESCENTS AND WE HAVE A VERY BROAD RANGE OF INDIVIDUALS WITH DIFFERENT PHYSIOLOGY MAYBE THIS IS A GREAT WAY TO DO TRIALS IN PATIENTS BUT BLEND. [INAUDIBLE] FROM THE TRIALS. >> YOU'RE ABSOLUTELY RIGHT. IN FACT, IN FACT I DID NOT MENTION THIS BECAUSE WE HAVE LIMITED TIME BUT EACH ONE OF THESE COULD BE A MAJOR TOPIC OF DISCUSSION BUT WITH THE CHP ONE OF THE MAJOR ADVANTAGES IS WE ARE CREATING THESE MODELS. THESE MODELS THAT YOU NOW GO AND GO AFTER. THOSE DECEASED MODELS ARE REALLY VERY GOOD BECAUSE IT'S NOT JUST THE TOXICITY OF THE DRUG, BUT IT'S ALSO GOING TO BE ITS EFFICACY. WE ALSO HAVE A SMALL MOLECULE PROGRAM HERE AT THE INSTITUTE'S WHERE WE ARE DEVELOPING SCOPE SMALL MOLECULES. I CAN TELL YOU THAT WE ARE DEVELOPING THESE CEASE MODEL IN THE SHIP THEN WE TEST THE EFFICACY OF THIS MOMENT YOU COOL MOLECULE AND THE CHIP. YOU CAN DO THAT BASED ON AGE RANGE TWO. CAN ACTUALLY GET ANOTHER AREA FOR EXAMPLE IS A TUMOR. WE'VE CURRENTLY LIKE 12 CLINICAL TRIALS AT LAKE FOREST FOR PATIENTS WITH CANCER. THAT INCLUDES BOTH PEDIATRIC AND ADULT BY THE WAY. WE CAN GET. [INAUDIBLE] A TUMOR BIOPSY AND WE CREATE A TUMOR AND A CHIP SO WE BUILD A CORE WITHIN THE INSTITUTE THAT HAS DEDICATED THIS JUST TO TUMORS. WE CAN THEN TEST THE BEST THERAPY ON THE TUMOR SO WE CAN PREDICT WITH THE BEST TREATMENTS CAN BE FOR THAT PATIENT USING THE CHEMOTHERAPY AGENTS. AS YOU KNOW YOU HAVE ALL THESE CHOICES AND CHEMOTHERAPY. WHAT CAN BE BEST FOR THAT PATIENT? THAT'S PERSONALIZED MEDICINE AT ITS BEST BECAUSE YOU'RE TAKING THE TUMOR FROM THE PATIENT AND THEN YOU'RE ACTUALLY GROWING A TUMOR AND A CHIP AND WE ARE NOW SEEING WHAT THE BEST TREATMENT IS FOR THAT SPECIFIC TUMOR ON THAT SPECIFIC PATIENT. WE HAVE ABOUT 12 DIFFERENT TUMORS NOW THAT WE ARE TACKLING BOTH PEDIATRIC AND ADULT. BECAUSE GOING TO BE THE NEXT STEP FOR THIS FIELD. IT'S NOT JUST LOOKING AT TOXICITY BUT LOOKING AT SAFETY, NOT JUST SAFETY BUT ALSO EFFICACY USING MODELS THAT ARE RELIABLE AND REPRODUCING. WHAT ARE THE CHALLENGES, YOU MENTIONED THAT YOU HAVE A 72 -YEAR-OLD KIDNEY DO BIOPSY COMING IN TO FIX A PROBLEM OR THE CHALLENGES THERE? OF COURSE AGING CAN INTRODUCE TROUBLES. ARE YOU ABLE TO REJUVENATE THE TISSUE IN ORDER TO GET BACK AND I MAKING A SIMPLE OR SAMPLE OF THIS. THIS FANTASTIC ALSO AND APPROACH FOR THE OVARIES AND ALL THE OTHERS THAT YOU HAVE SHOWN US. JUST AN IDEA HOW WE CAN TAKE SOMETHING HELLO THESE ARE INJURED IN AGED AND BRING IT BACK TO A FUNCTIONAL RECOVERY? >> THAT'S A GREAT QUESTION. THAT REALLY BRINGS US TO THE CELLS WILL BE ISOLATE. BASICALLY, AS YOU KNOW THE CELLS THAT WE REALLY ISOLATE EVERY TIME OR THE COMMITTED REGENERATIVE CELLS. THAT IS THE KEY TO ISOLATE THOSE CELLS. THOSE COMMITTED CELLS ARE THERE WHEN YOU'RE SEVEN-YEARS OLD AND THERE WHEN YOU'RE SEVEN-YEARS OLD. YOU'RE STILL THERE. AND WE TAKE SOMETHING LIKE THE PANCREAS FOR EXAMPLE AND YOU TAKE TYPE ONE DIABETES, LET'S TAKE TYPE ONE DIABETES FOR THE PANCREAS BASICALLY FINALLY BURNS OUT. IT WAS ACTUALLY HAPPENING IN THE PANCREAS IS THE ISLET CELLS ARE BEING FORMED AND ARE BEING DESTROYED. THEY ARE BEING FORMED AND DESTROYED AND ARE BEING FORMED AND DESTROYED SO EVENTUALLY THIS ENDS UP BEING TOTALLY BASED FIBROTIC AND IS TUMOR SPACE BUT THERE ARE STILL COMMITTED CELLS THERE AND THE PROBLEM IS I THINK IT'S TOO SCARRED THE MORE SCAR THERE IS THE HARDER IT IS FOR YOU TO GET LARGE QUANTITIES OF THOSE CELLS THAT ARE STILL THERE. WITH THE GOOD NEWS IS EVEN IN A 70 -YEAR-OLD PATIENT WITH END-STAGE SCARRING, YOU CAN STILL GET ENOUGH COMMITTED CELLS TO EXPAND THEM. ANOTHER DIFFERENCE THOUGH BETWEEN A SEVEN -YEAR-OLD NORMAL AND THE 70 -YEAR-OLD DECEASED IS IT'S GOING TO TAKE YOU LONGER TO GROW THE CELLS ON THE 70 -YEAR-OLD AND IT'S GOING TO. IT'S GOING TO AN EVENT OF THE DAY IT'S NOT THAT DRASTIC OF A DIFFERENCE IT'S GONNA TAKE YOU MAYBE A COUPLE OF WEEKS MORE. REMEMBER THE CELL GROWTH IS EXPONENTIAL SO YOU'RE JUST TAKING AN EXTRA FEW WEEKS BECAUSE THE NUMBER AND THE DENSITY OF YOUR CELLS IS LESS AND A SEVEN -YEAR-OLD THEN IN A SEVEN -YEAR-OLD BUT IT'S STILL THE SAME GROWTH POTENTIAL. >> LET'S LOOK AT THIS AND THIS IS PERSONALIZED MEDICINE AND EVERYTHING IS SHIPPING AND THE POTENTIAL ON THE OTHER HAND IF THE REGULATORY STANDPOINT THIS INTRODUCES SOME CHALLENGES BECAUSE BY AND LARGE THIS IS AND I GUESS YOU ARE TAKING YOUR SENSE AND MAYBE THERE IS ROOM FOR THIS BUT THIS IS GONNA BE THE CHALLENGE AND HOW DO YOU TACKLE THAT? >> WE BUILD THOSE AND THAT IS WHERE YOU BUILD THAT RELEASE CRITERIA AND THE RELEASE CRITERIA ARE SO IMPORTANT AS I MENTIONED BEFORE BUT REALLY WHEN YOU'RE LOOKING AT THE PATIENT TO PATIENT VARIABILITY THAT IS WHY IT'S IMPORTANT, I AM SO GLAD THAT YOU ASKED OF THIS BECAUSE REMEMBER, WE ARE ACTUALLY USING NORMAL HUMAN TISSUE AT FIRST. THIS IS TO FIGURE OUT A RELEASE CRITERIA THAT'S WHAT WE ARE DOING. WERE ACTUALLY TAKING BIOPSIES FROM YOUNG, OLD, MALE AND FEMALE DEPENDING ON THE ORGAN OF COURSE. SOME ARE ONLY MALE IN SUMMER ONLY FEMALE. BUT WHAT WE ARE TRYING TO DO THIS AND GOING TO THE CELL BIOLOGY STAGE, WE ARE TAKING A LARGE NUMBER OF BIOPSIES OF ALL KINDS OF PATIENTS YOUNG, OLD, NORMAL, DECEASED AND WHAT WE ARE SEEKING IS WE ARE SEEKING THAT RELEASE CRITERIA THAT'S GOING TO BE RIGHT IN THE MIDDLE. THAT IS VERY IMPORTANT BECAUSE IF YOU DON'T DO THAT, IF YOU DON'T DO THAT AND YOU END UP IN YOUR PATIENT PATIENT POPULATION AND LET'S SAY THAT YOU GET YOUR BIOPSIES WERE NORMAL PATIENTS AND YOU END UP IN THE PATIENT POPULATION THAT HAS A SPECIFIC DISEASE, YOU'RE NEVER GOING TO ACHIEVE YOUR RELEASE CRITERIA. THOSE BRACKETS THAT YOU ARE BUILDING ARE EXTREMELY IMPORTANT AND YOU ARE DOING THAT WITH HUMAN TISSUE BIOPSIES THAT YOU ARE DOING TIME AND TIME AGAIN TO MAKE SURE YOU CAN GET THAT TO RELEASE CRITERIA THAT WILL BE BROAD ENOUGH FOR THE WHOLE POPULATION. >> IS PROBABLY ENOUGH TO GIVE YOU LATITUDE BUT IT ALSO GIVES A RANGE SO IF YOU LOOK AT IT THE SOLUTION AND YOU CAN TRY TO BE WITH THE MAJORITY AND WHERE THE PATIENTS ARE. I CAN GO ON AND ON AND ON AND REALLY WE WANT ANOTHER PATIENT AND THEN WE LEAVE IT THERE BECAUSE WE WANT DON'T WANT TO ABUSE YOUR TIME. THIS IS RELATED TO THE EGGS AND HOW TO ENGINEER THE OVARY SO HOW MUCH DATA DO YOU THINK WILL BE NECESSARY TO GET TO THE DOOR OF THE FDA INTO GET TO A KNOCK AT THE DOOR? I'M SIMPLIFYING THIS I KNOW. >> THERE IS A LONG ROAD AHEAD. IT'S TAKEN YEARS FOR US TO GET TO THIS POINT I TELL YOU. IT'S A LONG-STANDING PROJECT FOR US ABOUT GETTING OVARIAN CELLS AND STEM CELLS. WHEN YOU LOOK AT A POSTMENOPAUSAL TISSUE THERE ARE NUMBER FOLLICLES THAT CAN BE RETRIEVED THERE ARE NO FECAL SALES, NO GRANULAR SOCIAL CELLS THAT SUPPORT NO POLAR BODIES AND RELYING ON A STEM CELL POPULATION WITHIN THE TISSUE LUCKILY THEY'RE STILL THERE AND LIKE WE TALKED ABOUT, EVEN A SEVEN -YEAR-OLD HAS THOSE COMMITTED REGENERATIVE CELLS. IT'S BEEN A LONG ROAD TO TRY TO GET THOSE CELLS TO GET THE STEM CELL POPULATIONS TO FORM. STEM CELL POPULATIONS TO FORM GRANULE CELLS AND THOSE TO FORM SITES TO PUT THE WHOLE THING AND ACTUALLY ENGINEER THE FOLLICLE SO THAT YOU HAVE ALL OF THOSE LAYERS. IT HAS TAKEN A LONG TIME TO ENGINEER THIS AND TO THE POINT WHERE WE ARE TODAY WE HAVE THIS AND IT'S REALLY GREAT STEM CELLS. WE ARE NOW GOING TO START DOING THE INTRA- CYTOPLASMIC SPERM INSERTION. AND SO THE MAJOR QUESTION FOR US IS HOW NORMAL ARE THESE EGGS? HOW NORMAL ARE THEY REALLY? WE HAVE BEEN CHARACTERIZING THESE AS WE MAKE THEM TO MAKE SURE THEY ARE IN THE NORMAL RANGE BUT WILL THEY HAVE THE RIGHT INTEGRITY FOR SPERM TO SURVIVE? WE DON'T KNOW OR JUST STARTING THOSE EXPERIMENTS WAS GOING TO HAPPEN WITH THOSE? SO MANY QUESTIONS TO BE ANSWERED AND THIS IS STILL WORK AND DEVELOPMENT AND IT'S GOING TO BE MARRIED YEARS BEFORE THIS GETS TO THIS PATIENT. >> WE CAN STOP HERE AND AGAIN THANK YOU FOR YOUR FANTASTIC LEANING THE FIELD FOR YOUR ABILITY TO SHARE THAT VERY SHORT TIME THE SPECTRUM OF WHAT YOU HAVE BEEN DOING AND WHERE YOU SEE THIS MOVIE AND FOR SHARING YOUR INSIGHTS TODAY. THIS IS AGAIN STARTING A CONVERSATION FOR US AND WE WOULD LIKE TO CONTINUE THIS IDEA ESCAPE. THE MOVE INTO BREAK NOW, WE WANT TO THANK YOU ON BEHALF OF THE NH ID IN THE COMMUNITY FOR THE VALUABLE INSIGHTS THAT YOU SHARED WITH US TODAY. WE WILL BE IN TOUCH I'M SURE. >> THANK YOU SO MUCH AS SCRAPING WITH ALL OF YOU TODAY THANK YOU. WE WILL MOVE ON TO A BREAK AND WE WILL BE JUST A 12 MINUTE BREAK AGAIN I WOULD ASK THE LOGISTICS TEAM TO PLEASE PUT THE SLIDES ON SO WE HAVE THAT LINK TO THE IDEAS AND WE WILL GO TO 240 FOR THE NEXT SESSION THANK YOU VERY MUCH FOR THIS ENGAGING CONVERSATION THIS MORNING. WELCOME BACK. WE'RE HEADING TOWARDS THE NEXT STATION. SESSION 4. WE HAVE AS A MODERATOR OUR OWN -- FROM THE -- OF THE NICHD WHO IS MODERATING THIS SESSION AND I WILL LET HER TAKE IT AWAY >> HI. GOOD AFTERNOON, EVERYONE. MY NAMING CLARA CHEN AND I'M A PROGRAM OFFICER AT THE FERTILITY AND -- BRANCH NICHD. IT'S AN HONOR TO BE THE MODERATOR OF THIS SESSION AND A PRIVILEGE TO INTRODUCE OUR PRESTIGIOUS SPEAKERS. BEFORE I INTRODUCE OUR FIRST SPEAKER I WOULD LIKE TO REMIND YOU THAT DURING THE PRESENTATION IF YOU HAVE ANY QUESTIONS YOU CAN SUBMIT YOUR QUESTIONS THROUGH THE LIVE FEEDBACK AT THE BOTTOM OF THE VIDEO CAST WEBSITE. AND SO OUR SPEAKER CAN ADDRESS YOUR QUESTION AFTER THE TALK OR DURING THE PANEL DISCUSSION AND IF YOU'RE ONE OF THE PANELIST THEN YOU CAN PUT YOUR QUESTIONS IN THE ZOOM CHAT. OKAY. SO OUR FIRST SPEAKER IS DR. JENNIFER LEWIS. -- I BELIEVE YOU ALL KNOW HER. SHE IS A RENOWNED RESEARCHER AND ALSO A NATION WILD LEADER IN THIS FIELD BUT I STILL TRY TO DO MY JOB DILIGENTLY. SO A BRIEF INTRODUCTION FOR DR. LEWIS. A FULL PROFESSOR AT HARVARD THE UNIVERSITY. FOCUS US ARING ON 3-D PRINTING OF FUNCTIONAL AND STRUCTURAL AND BIOLOGICAL MATERIALS. CURRENTLY DR. LEWIS DIRECTS THE RESEARCH SCIENCE AND ENGINEERING CENTERS AND SERVICES ON THE -- -- OF THE ADVISORY COMMITTEE. DR. LEWIS HAS RECEIVED NUMEROUS AWARDS INCLUDING THE PRESIDENTIAL FACULTY FELLOW AWARD AND SHE IS AN ELECTED MEMBER OF THE NATIONAL ACADEMY OF SCIENCE NATIONAL ACADEMY OF ENGINEERING, NATIONAL ACADEMY OF INVENTORS AND AMERICANA ACADEMY OF ARTS AND SCIENCE. SHE HAS CO-FOUNDED MULTIPLE COMPANIES THAT ARE COMMERCIALIZING TECHNOLOGY FROM HER LABORATORY. THE TITLE DR. LEWIS TALK IS BIOMANUFACTURING, VASCULARIZED HUMAN TISSUE FOR REPAIR AND REGENERATION. PLEASE TAKE IT AWAY DR. LEWIS. >> THANK YOU ALL. I WOULD LIKE TO SHARE MY SCREEN. THANKS. AS CLARA MENTIONED TODAY I'M GOING TO TALK ABOUT SOME OF THE WORK WE'VE BEEN DOING IN BIOMANUFACTURING VASS KULAR RISED HUMAN TISSUES. THESE IS ALSO RELEVANT FOR DRUG TESTING AND DISEASE MODELING. IF WE THINK ABOUT THESE APPLICATIONS -- THAT I JUST DESCRIBED THERE IS REALLY AN URGENT NEED FOR RENEWABLE SOWS SOURCES OF HUMAN TISSUES. FOR BOTH * CARRYING OUT A PHASE 0 FAIL FAST TRIAL ON DRUG TOXICITY TO DISEASE MODELING TO ADDRESSING SOME OF THE KEY SHORTAGES AND CHALLENGES IN ORGAN ENGINEERING. SO IF WE THINK ABOUT PATIENTS AWAITING ORGANS WORLDWIDE THE KIDNEY, THE HEART, THE LIVER ARE SOME OF THE MOST IMPORTANT ORGANS OF NEED. AND IF ONE COULD START WITH PATIENT SPECIFIC CELLS AS THE SOURCE OF THE MATERIAL ONE COULD ACTUALLY MAKE AN ORGAN THAT IS MADE FROM YOU FOR YOU. AND THAT WOULD REDUCE REJECTION AFTER TRANSPLANTATION AND ADDRESS THIS EVER WIDENING GAP. OUR WORK -- STARTED IN THIS FIELD PROBABLY 10 YEARS AGO WHEN WE MADE AN IMPORTANT DISCOVERY. AND THAT IS THE IDEA OF USING A SACRIFICIAL MATERIAL. INTO DENSE THREE DIMENSIONAL TISSUES. YOU'RE SEEING A BONE TYPE IMPLANT THAT IS BEING PATTERNED. THESE POSTS THAT ARE COMING UP ARE ACTUALLY THE SACRIFICIAL INK AND IT'S PENETRATING THIS TISSUE THAT IS ABOUT A CENTIMETER THICK AND THAT ALLOWS US TO DELIVER GROWTH FACTORS AND DIFFERENTIATE STEM CELLS IN PLACE AND ALSO SUPPLY NUTRIENTS TO THIS ISSUE. AT THE SAME TIME WE INVENTED ANOTHER METHOD CALLED EMBEDDED 3-D BIOPRINTING. THIS IS AN EMBEDDED PRINTING METHOD WHERE WE CAN GO DOWN INSIDE A TISSUE MATRIX. A CELLULAR MATRIX. 8 BUT YOU CAN WRITE A COMPLEX VASCULAR TREE AND ULTIMATELY WE CAN ERASE THAT INK SO WHERE YOU'RE SEEING THE PINK INK HERE ULTIMATELY WE CAN TAKE THIS TISSUE AFTER WE'VE LAID DOWN THESE VASCULAR CONDUETS AND OPEN UP THE CHANNELS AND PERFECT FUSE THE TISSUE IN BULK AND SUSTAIN THE CELTS. PRINTING CELL LADEN INKS -- EVEN WITH THE CAPABILITY -- DOESN'T REALLY MEET THE NEED FOR ENGINEERING SOLID ORGANS AS ANTHONY DESCRIBED EARLIER. IF WE WANT TO ADDRESS THE KIND OF COMPLEX FUNCTIONS AND SCALE THAT ONE SEES IN A HEART AND KIDNEY AND LIVER WE HAVE TO BE ABLE TO PATTERN TISSUES AT HUNDRED OF BILLIONS OF CELLS AND HIGH CELLULAR DENSITIES AND MOST CONTAIN MULTIPLE CELL TYPES PER ORGAN AND THEY ARE ARRANGED IN A COMPLEX ARCHITECTURE. THAT I WOULD ARGUE IS DIFFICULT TO PRINT FROM THE TOP DOWN. 7 SO IN ORDER TO SPAN THESE MULTIPLE SCALES AND MULTIPLE CELL TYPES AND YET STILL BE ABLE TO DO THIS IN A SCALABLE WAY WE HAVE PIVOTED TOWARDS A HYBRID BIOPRINTING/SELF-ASSEMBLY APPROACH AND THE IDEA IS TO REALLY LEVERAGE SOME OF THE MANY ADVANCES THAT HAVE OCCURRED OVER THE LAST DECADE PLUS FORGETTING RENEWABLE CELL SOURCES DIRECTLY FROM PATIENTS AND BEING ABLE TO ASSEMBLE THESE INTO ORGAN BUILDING BLOCKS. THESE COULD BE ORGANOIDS THAT HAVE BEEN DIFFERENTIATED SUCH AS THE KIDNEY ORGANOID. BUT THE IDEA IS TO TAKE THESE SMALL BUILDING BLOCKS AND USE THOSE AS THE BIOMATERIAL. LET'S NOT PRINT THE SCAFFOLD -- LET'S PRINT THE CELLS THEMSELVES OR LET'S ASSEMBLE THESE CELLS INTO A MATRIX IN WHICH WE CAN PRINT VASCULAR. WHAT YOU'RE SEEING HERE IS THE SAME IDEA OF WRITING A SACRIFICIAL LINK INTO THE TISSUE AND NOW THE TISSUE IS COMPRISED OF THESE ORGAN BUILDING BLOCKS. AND THESE ARE MULTI-CELLULAR AGGREGATES THAT HAVE BEEN DERIVED FROM ISPCs AND EACH ONE IS BEATING ASYNCHRONOUSLY IN THIS CASE. WE'RE USING THIS CONCEPT OF BIOPRINTING WITH BIOLOGICAL SELF-ASSEMBLY TO CREATE A MYRIAD OF TISSUE TYPES START WITH ORGAN BUILDING BLOCKS SUCH AS THESE KIDNEY ORGANOIDS ALL THE WAY TO THE CARDIAC -- AND ACROSS THIS ENTIRE SPACE BECAUSE THERE ARE SO MANY DIFFERENT PROTOCOLS EMERGING FOR ORGANOIDS AND MULTI-CELLULAR AGGREGATES FROM ISPCs WE HAVE THE ABILITY TO CREATE THESE TISSUES IN A VARIETY OF SIZES AND SCALES WITH EXQUICKIC ARCHITECTURE * SUCH AS SHOWN HERE. AND PROXIMAL TUBULAR SEGMENTS AND VASCULAR THAT PERVADES THE ORGAN BUILDING BLOCKS AND THEN COUPLED WITH THAT WE CAN PRINT AT A LARGER SCALE USING THESE SACRIFICIAL INKS A MACRO VESSEL NETWORK SO BY COMBINING THESE APPROACHES WE BELIEVE WE CAN ADDRESS SOME OF THE KEY CHALLENGES WITH THE DESIRE FOR HIGH CELLULAR DENSITY, HIGH NUMBER OF CELL PIPES AND THE ABILITY TO SCALE IN THREE DIMENSIONS. SO I WANT TO HIGHLIGHT TWO PEOPLE THAT HAVE REALLY BEEN FOUNDATIONAL IN THIS WORK THAT WE CALL SWIFT. WHICH IS SACRIFICIAL WRITING INTO FUNCTIONAL TISSUE. THE IDEA HERE IS I'VE SHOWN -- ALREADY IS WE START WITH IPSCs AND CREATE THESE AGGREGATES. IN SOME CASES THEY ARE ORGANOIDS AND OTHER CASES THEY HAVE NOT BEEN DIFFERENTIATED. WE CREATE THIS EMBRYO OF BODIES AND THEN WE PATTERN THIS SACRIFICIAL INK AND REMOVE THE INK LEAVING BEHIND THE PERVASIVE VASCULAR THAT CAN BE PROFUSED. SO ONCE WE HAVE PATTERNED THIS THE NEXT STEP IS TO TAKE THAT TISSUE. WE ALREADY HAVE IT EMBEDDED IN A BIOREACTOR AND WE CAN PROFUSE THOSE CHANNELS TO SUSTAIN AND MATURE THIS TISSUE AND THESE INDIVIDUAL ORGAN BUILDING BLOCKS ARE GOING TO FUSE TOGETHER TO CREATE A CONTIGUOUS THREE DIMENSIONAL TISSUE AND OUR GOAL IS TO CREATE A FUNCTIONAL TISSUE. SO WE HAVE TO CHARACTERIZE THE FUNCTIONAL OUTPUT OF THE TISSUE. WHAT YOU'RE SEEING HERE IS A MOVIE. IN THIS MOVIE WE HAVE ESSENTIALLY 400,000 'E EMBRYO BODIES. 7 THIS IS A NOZZLE TRANSLATING THROUGH NOT PRINTING. NOW WE'LL SEE IT COME BACK THROUGH AND PRINT THE SACRIFICIAL INK. THIS ALLOWS IT TO MOVE THROUGH EASILY. THIS TISSUE HAS ROUGHLY HALF A BILLION CELTS AND IT'S ROUGHLY TWO MILLILITERS IN VOLUME. SO WITH THIS TECHNIQUE THEN WE CAN PATTERN THE VASCULATURE THAT WE WANT AND IT HARKENS BACK TO THE ABILITY TO TAYLOR AND USE THE GRANULAR LIVING BUILDING BLOCKS WHERE WE GENERATE LARGE VOLUMES OF THIS. MILLILITERS WORTH OF TISSUE AND WE CAN ACTUALLY MEASURE THE REALITY OF THE LIVING MATRIX ITSELF AND THIS IS US LOADING AN A ASSEMBLY OF THESE LIVING TISSUE BUILDING BLOCKS INTO THE RE -OMETER AND ALSO THEY HAVE A -- THAT ALLOWS US TO PATTERN THE VASCULATURE WITHIN THEM AND RETAIN THAT VASCULATURE IN PLACE UNTIL WE REPLACE IT. SO THIS LIVING MATRIX REALGAE IS IMPORTANT. *. THESE ARE THE ORGAN BUILDING BLOCKS THEMSELVES. WE HAVE A SMALL PERCENTAGE OF EXTRA CELLULAR MATRIX THAT WE SPECIFICALLY DESIGNED TO BE IN ACTIVE DURING THE PRINTING PROCESS NOT CONTRIBUTING TO THE REALITY OF THE SYSTEM BUT WHEN WE WARM UP THE TISSUE TO 37 DEGREES C THE EXTRA CELLULAR MATRIX GEL HOLDS THE AGGREGATES IN PLACE WHEREAS THE SACRIFICIAL INK MELTS AT THAT TEMPERATURE ALLOWING US TO FLUSH AWAY THE PLACES WHERE WE HAVE EMBEDDED THESE CON DUE LIMITS INSIDE THE TISSUE. *. WHAT YOU SEE ARE THE OPEN LOOM ENS. AND COLLAGEN ONE WHICH IS PART OF THE MATRIX THAT WE USE HERE TO FORM THIS STABLE MATRIX THAT WE CAN ERASE AWAY THE INK FROM. SO LET'S TAKE ANOTHER PEEK AT THIS. THIS PROCESS YOU CAN SEE WE'VE GOT A LARGE ASSEMBLY OF ORGAN BUILDING BLOCKS. WE'VE GOT AN INLET AND OUTLET IN THIS DEMONSTRATION AND WE'VE PATTERNED A PERVASIVE VASS COOLA YOU ARE. JUST BY CHANGING * THE SPEED AT WHICH WE TRANSLATE THE NOZZLE AT AFFIXED DEPOSITION RATE WE CAN CREATE CON DUETS THAT ARE * LARGER OR SMALLER IN SIZE AS YOU CAN SEE HERE AS A CROSS -- CROSS SECTIONAL CUT. MUCH OF WHICH ARE ALIVE IF WE LOOK AT A LIVE DEAD STAIN. WE CAN TAYLOR THE LUMEN SIZE USING A SINGLE NOZZLE BY CHANCING THE TRANSLATION SPEED. IF WE DON'T EMBED THAT VASCULATURE OF COURSE WE'RE GOING TO DEVELOP A NECROTIC CORE. SO THIS ABILITY TO SACRIFICIALLY WRITE THE CHANNELS WITHIN THE SYSTEM IS CRUCIAL. AND AS I'VE SHOWN HERE IF WE CAN DO THIS SUCCESSFULLY THEN WE CAN SUSTAIN QUITE A HIGH CELL VIABILITY EVEN IN TISSUES THAT ARE AS DENSE AS ABOUT 200 CELLS PER MILLILITER. SO LET ME JUST GIVE YOU ONE EXAMPLE OF WHAT WE'RE DOING WITH THIS SWIFT AND THIS IS TO CREATE CARDIAC TISSUING CARDIAC ORGAN BUILDING BLOCKS. WE'RE FOLLOWING A PROTOCOL -- -- TO BASICALLY SCALE AND GENERATE THESE BUILDING BLOCKS. WE START WITH ISPCs. WE GROW THEM UP. USE A MICRO WELL OF REYES TO FORM INDIVIDUAL SPEAR RIDES. * AND THOSE ARE THE A MIXTURE OF CARDIAC MAYO CITES AND A SMALL -- OF POPULATION. THESE BUILDING BLOCKS HAVE HIGH CELLULAR DENSITIES EXCEEDING 200 MILLION CARDIO MAYO SITES PER * INDIVIDUAL AGGREGATE. AND AS I SHOWED EARLIER THEY POST NINE DAYS AFTER DIFFERENTIATION EACH ONE OF THESE BECAUSE THEY ARE ISPC DERIVE BEAT ASYNCHRONOUSLY. WE CAN COMPACT AND WRITE THE VESSELS INSIDE. SO THIS IS A MOVIE OF THAT. WHERE WE HAVE A PIN DOWN BELOW AND CONNECTING UP HERE THIS IS JUST A SINGLE BRANCHING ARCHITECTURE AFTER WE ERASED THE INK WE'RE JUST SHOWING THAT WE CAN PERFECT FUSE A DYE * THROUGH THAT AND YOU CAN SEE THE OPEN CHANNELS WITH A MEDIA THAT IS DYED BLUE SO WE CAN SHOW YOU THAT WE CAN PERFECT FUSE THROUGH THE TISSUE. THIS IS A CROSS SECTIONAL. THERE IS A SINGLE VESSEL BEING PATTERNED AND THIS SHOWS A HIGHER MAGNIFICATION VIEW OF THE MACHINERY THAT IS RESPONSIBLE FOR THAT CONTRACTION OF THE HEART TISSUE. SO ON DAY ONE IF WE LOOK AT THIS TISSUE NOW THAT IT'S MORE COMPACTED THESE INDIVIDUAL SPHERES ARE MORE DENSELY PACKED THEY ARE STILL BEATING ASYNCHRONOUSLY. AT DAY 10 IT HAS FUSED. AND NOW WE HAVE A TISSUE THAT BEATS SYNCHRONOUSLY. AS I SAID THESE TISSUES COULD BE USED FOR DRUG TESTING. THIS IS AN EXAMPLE THAT SHOWS IF WE SUBJECT THIS TISSUE BY PERFECT FUSING CONCENTRATIONS OF KNOWN DRUGS THAT EITHER SPEED UP THE BEAT FREQUENCY OR SLOW DOWN. -- AND IF WE LOOK AT THE SYNCHRONOUS BEATING ON DAY ONE WE HAVE THESE INDIVIDUAL BUILDING BLOCKS THAT ARE ALL DOING THEIR OWN THING BUT BY DAY SEVEN AFTER TISSUE FUSION HAS OCCURRED WE GET A NICE SYNCHRONOUS BEATING AND THIS OCCURS SPONTANEOUSLY. WE CAN LOOK AT TWO DIFFERENT REGIONS HERE ON THE LEFT AND RIGHT IN BOTH OF THESE IMAGES AND WE CAN COMPARE THOSE TO ONE ANOTHER. ON THE LEFT AND THE RIGHT IF YOU WILL. BOTH AT DAY ONE AND DAY SEVEN WHEN THEY ARE SPONTANEOUSLY BEATING OR WHEN -- AND OF COURSE WE HAVE THIS A SYNCHRONOUSY THAT -- AND IT LOOKS VERY NICE AS IT IS BEING PACED ELECTRICALLY. HOW DO WE GO FROM JUST PATTERNING THE LARGER CHANNELS AND VESSELS TO GETTING VASCULATURE AT THE MICRO CAPILLARY LEVEL WITHIN EACH OF THESE BUILDING BLOCKS? ONE APPROACH IS TO SIMPLY ADD -- CELLS TO THESE MULTI-CELLULAR AGGREGATES AND WE'VE DONE THAT RECENTLY AND WE SEE THAT LEADS TO QUITE A HIGHER DEGREE OF CONTRACT -- THE FUNCTIONAL OUTPUT OF THE TISSUE IS ENHANCED. WE SEE A NETWORK WITHIN THESE BUILDING BLOCKS AND YOU CAN SEE BEAUTIFUL ORGANIZATION OF THE -- THAT ARE RESPONSIBLE FOR THIS TISSUE CONTRACTION. AND ALSO OF COURSE WE ULTIMATELY WANT TO BE ABLE TO SEE THESE CHANNELS. WITH ENDO FILLIAL CELLS. *. WE TAKE THAT AND WE CAN INTRODUCE THE CELLS TO LINE THOSE CHANNELS AND ULTIMATELY PROMOTE INTEGRATION BETWEEN THE MICRO VES -- VESSELS. I HOPE THIS GIVES YOU A SENSE OF WHERE WE WANT TO GO WITH THIS WORK. USE SWIFT TO RECONSTRUCT TISSUES FROM PATIENT SPECIFIC CT DATA. THIS IS AVAILABLE ON THE NIH 3-D PRINT WEBSITE AND WE'VE TAKEN THAT TISSUE WEDGE OUT OF THIS VENTRICLE AND WE'VE PATTERNED AND EMBEDDED IN HERE THE SEPTIL AND DISTAL BRANCHES AND YOU CAN SEE THAT THERE. SO JUST TO REMIND YOU IN CLOSING THAT WE BELIEVE THIS APPROACH WILL WORK WITH ANY ORGAN BUILDING BLOCK AND WE'VE BEEN WORKING WITH BOTH CARDIAC AS WELL AS KIDNEY ORGAN BUILDING BLOCKS TO CREATE KIDNEY TISSUE BUT IT SHOULD BE AMENABLE TO ANY BUILD'ING BLOCK THAT IS MULTI-CELLULAR IN NATURE. WITH THAT I WILL END AND SAY THAT IN ADDITION TO THE CARDIAC AND KIDNEY WORK WE'VE ALSO BEEN WORKING WITH NEURAL TISSUES AND HERE WE'VE BEEN PREPROGRAMMING CELLS USING TRANSCRIPTIONAL FACTORS TO -- DEFINE SPECIFIC CELL POPULATIONS. SO WE START WITH THE SAME IPSC AS WILD TYPES AND THEN WE USE TRANSCRIPTION FACTORS TO PROGRAM THEM INTO VAS VASCULATURE OR NEURONS. THESE ARE VERY DENSELY CELLULAR STEM-CELL BASED INKS WITH THE GOAL OF PLACING TISSUE FUNCTION AT THE HEART OF THIS BIOMANUFACTURING APPROACH. SO WITH THAT I'LL END AND THANK THE TEAM AND SOME OF OUR KIKO LAB RATERS IN THE CARDIAC SPACE. WE'RE WORKING WITH CHRIS CHEN. FROM THE PERSPECTIVE OUR KIDNEY -- INITIATIVE - --- I WOULD LIKE TO THANK THE FUNDING THAT I'VE RECEIVED. WITH THAT I'LL END AND TAKE ANY QUESTIONS THAT YOU MAY HAVE. >> THANK YOU, DR. LEWIS. SUCH A FASCINATING WORK AND EXCELLENT PRESENTATION. SO IF YOU DON'T MIND WE -- >> WE'RE GOING TO HAVE A PANEL DISCUSSION. LET ME JUST STOP SHARING. THANK YOU. >> THANK YOU. >> I WOULD LIKE TO INTRODUCE OUR NEXT SPEAKER. 7 DR. -- -- IT'S YOUR TURN. YOU CAN START TO UPLOAD YOUR SLIDE. SO DR. -- IS AN ASSOCIATE PROFESSOR IN THE DEPARTMENT OF BIOENGINEER AT THE UNIVERSITY OF ILLINOIS AT CHICAGO. DR. -- HAS OVER 20 YEARS EXPERIENCE IN THE BIOMATERIALS AND TISSUE ENGINEERING FIELDS. HER WORK RELATED TO EVERYTHING FROM OVER -- ORGAN AND MUSCULAR SKELETAL TISSUE BY FABRICATION TO NEW METHODS FOR PRINTING METALS. HER WORK HAS LED TO OVER A DOZEN ISSUES AND PENDING PATENTS. AND ALSO -- HAS IMPACT -- SHE HAS BEEN FEATURED -- AND CHICAGO BUSINESS MAGAZINE AND ALSO FOX NEWS WHEREAS OTHER NATIONAL AND INTERNATIONAL MEDIA OUTLETS. DR. -- PLEASE TAKE IT AWAY. >> THANK YOU FOR THE IN TREMENDOUS DUCKS AND THE OPPORTUNITY TO PARTICIPATE. TODAY I'M TALKING ABOUT SOME OF THE WORK WE'VE BEEN DOING OVER THE YEARS AND TRYING TO CREATE ARTIFICIAL MICRO ENVIRONMENTAL. I'M THE CO-FOUNDER AND CSO OF -- INKS. BY GET INTO THE DEPTH OF SOME OF THE WORK THAT WE'VE BEEN DOING I WOULD LOVE TO FIRST ACKNOWLEDGE THE TEAM WHO'S HERE. THEY WERE THE ORIGINAL 3-D PRINTED ARTIFICIAL TEAM AT NORTHWEST UNIVERSITY STARTING OFF IN 2013 WITH MY COLLABORATION WITH DR. TERESA WOODRUFF. AND HER TEAM BACK THEN -- PARTICULARLY DR. MONICA LARONDA -- AND WHO I'M CONTINUING THIS WORK AFTER THESE INITIAL STUDIES I'M PRESENTING. DR. ALEX RUTZ WAS THE GRADUATE STUDENT ON MY TEAM WHO DID THE DEVELOPMENTS OF THE MATERIALS. SO THESE ARE THE FOUNDING TEAM THAT LED TO THE WORK THAT I'M PRESENTING HERE. AND THAT REALLY STARTED OFF THIS JOURNEY. SO JUST STARTING OFF WITH WHY ARE WE FOCUSING ON TRYING TO CREATE ARTIFICIAL OVARY. ADDRESS THE CURRENT LIMITATIONS ON OPTIONS THAT ARE HERE TODAY. THESE ARE FOR PATIENTS WHO WANT TO POSTPONE PREGNANCY, THOSE WHO ARE CANCER PATIENTS WHO CANNOT UNDERGO FERTILITY TREATMENTS. AT THE PRESENT BECAUSE OF DIFFERENT CANCER TREATMENTS. AND SO THE CURRENT GOLD STANDARDS FOR PRESERVING FERTILITY IS EGG OR EMBRYO BANKING. WOMEN HAVE TO UNDERGO MANY INJECTIONS OF HORMONE THERAPIES TO STIMULATE THEIR FOLLICLES TO GENERATE MATURE EGGS INSIDE THEIR BODY. AFTER THEY MATURE THESE EGGS CAN BE RETRIEVED AND EITHER BE FROZEN OR GO THROUGH THE PROCESS AND BANKED FOR FUTURE USE. THE THING ABOUT THIS STRATEGY IS IT DOES TAKE TIME TO ACTUALLY MATURE EGGS IN THE WOMEN'S BODY AND SOMETIMES IT TAKES MANY CYCLES. SO FOR THOSE WHO HAVE CANCER ESPECIALLY AGGRESSIVE CANCER THIS MAY NOT BE AN OPTION BECAUSE THEY NEED TO HAVE CANCER TREATMENTS RIGHT AWAY. ALSO MANY PATIENTS WHO HAVE UNDERLYING MEDICAL CONDITIONS THAT DON'T RESPOND TO HORMONE STIMULATION. AND SO IF WE CAN FIND A WAY TO CREATE ARTIFICIAL ENVIRONMENTS TO BE ABLE TO ATTAIN THE FOLLICLES AND CULTURE THEM AND MATURE THEM OUTSIDE THE BODY INSTEAD OF WITHIN THE PATIENT THAT WOULD BE ANOTHER OPTION WE CAN PROVIDE USING THESE TISSUE ENGINEERING STRATEGIES. IMPORTANTLY THESE HORMONE STIMULATION TREATMENTS CANNOT BE APPLIED TO -- FERTILE PATIENTS PARTICULARLY CANCER PATIENTS. THERE HAS BEEN GREAT PROGRESS AS FAR AS CANCER TREATMENTS BUT THERE IS A SIGNIFICANT CORRELATION BETWEEN THE RADIATION THERAPY AND DOWNSTREAM INFERTILITY -- OVARIAN FAILURE THROUGHOUT THEIR LIVES AND THESE LOWER THAN NORMAL HORMONE LEVELS CAN ALSO LEAD TO OTHER MEDICAL PROBLEMS SUCH AS OSTEOPOROSIS OR CARDIAC ISSUES AND SO ONE MORE RECENT I GUESS RELATIVELY RECENT STRATEGY IS TO AND THIS IS A SCHEMATIC OF AN OVARY. WE CAN TAKE A PIECE OF THE CORTEX TISSUE OF THE OVARIES. THIS OUTER TISSUE THAT WITHIN IT PRESIDES FOLLICLES SO THE EARLY STAGE FOLLICLES HAS THE -- -- IT HAS THESE GRANULAR CELLS AND HORMONE PRODUCING CELLS THAT MAINTAIN VIABILITY AND FUNCTION. THEY TAKE THESE FOLLICLES WHICH ARE MOST IN THE CORTEX. THEY TRY TO PRESERVE IT AND AFTER TREATMENT OR AS THE PATIENT GROWS OLDER THEY ARE THEN ABLE TO TRANSPLANT IT BACK INTO THE PATIENT UPON WHICH THEY CAN REGAIN IN THEIR FERTILITY AND NORMAL HORMONE PRODUCTION. THIS HAS BEEN SUCCESSFUL AND THIS IS ABOUT 135 BIRTHS REPORTED WORLDWIDE. HOWEVER THE BIRTH RATE IS STILL RELATIVELY LOW AT 30% AND THE BIGGEST RISK IN THIS PROCEDURE IS THE POSSIBILITY OF REINTRODUCING CANCER CELLS BECAUSE THERE ARE STUDIES THAT HAVE SHOWN THAT CANCER CELLS CAN RESIDE WITHIN THIS TISSUE. SO IF WE'RE ABLE TO CREATE AN ALTERNATIVE WHERE WE CAN ISOLATE THE FOLLICLES AND HOUSE THEM OUTSIDE THE BODY AND MAINTAIN THE VIABILITY AND DOWNSTREAM IMPLANT THEM BACK INTO THE PATIENT THAT LOWERS THE RISK OF INTRODUCING CANCER CELLS. THAT WOULD BE A MORE IDEAL OPTION FOR THESE PATIENTS. SO QUICKLY AS FAR AS AND THIS HAS TOUCHED UPON THE TALK WE REALLY NEED TO UNDERSTAND WHAT WOULD IT TAKE -- TO CREATE A FUNCTIONING ARTIFICIAL ORGAN. IN THIS CASE THE OVARY AND THIS IS A DIFFERENT SCHEMATIC WHERE THE ARTIFICIAL ENVIRONMENT WILL BE PRESENTING TO THESE FOLLICLES HAS TO BE SUCH THAT IT SUPPORTS OR PROVIDES THE RIGHT CHARACTERISTICS AND SUPPORTS THE VIABILITY AND GROWTH. THIS IS AN EXAMPLE OF FOLLICLES THAT ARE CULTURED -- LIKE MANY OTHER CELLS THEY CHANGE AND IN THIS CASE THE FOLLICLES DIE AND THIS SHOWS THE DISASSOCIATION OF THE FOLLICLE -- WHERE THE CELLS MOVE AWAY FROM THE EGG CAUSING EGG DEATH. THIS IS MENTIONED BEFORE. REALLY IN OUR WORK WE REALLY STRIVE TO THINK ABOUT TRANSLATION AND SURGICAL IMPLANTATION IN CREATING DEVICES THAT CAN ACTUALLY BE EASILY IMPLEMENTED SURGICALLY. SO, THAT IS A BIG PART. ONCE IT IS IMPLANTED VASCULARIZATION IS IMPORTANT. BEING ABLE TO MAINTAIN VIABILITY OF THE CELLS BUT IN THE CASE OF OVARY IT HAS TO CIRCULATE THE HORMONES FROM THE FOLLICLES. IT HAS TO BE ABLE TO RELEASE THE EGGS -- FOR -- FERTILIZATION. WE'VE TRIED TO UNDERSTAND THE NATURAL ENVIRONMENT OF THE OVARY AND SHE HAS BEEN ABLE TO SHOW THERE ARE DIFFERENT REGIONS THAT THE OVARY THE OVARY ORGAN AND DIFFERENT FOLLICLES RESIDE IN DIFFERENT PLACES. AND WITHIN THOSE PLACES THERE ARE DIFFERENT DENSITIES AND MECHANICAL PROPERTIES. THE CORTICAL TISSUE -- -- UNTIL THERE IS A SIGNAL TO INDUCE ACTIVATION AND GROWTH. AND SO THEY RESIDE MORE IN THE CORTEX AND WHEN THEY ARE ACTIVATED AND THEY START TO GROW THEY HAVE TO MIGRATE DOWNSTREAM -- DOWN DIFFERENT PATHS WITHIN THE OVARY THAT IS A LESS -- AREA AND THE PROTEINS THAT ARE PRESENT WITHIN A DIFFERENT REGION OF THE OVARY SHOWING THERE IS MORE COLLAGEN ONE IN THE CORTEX AND LESS IN THE MEDULLA. ONE THING THAT INTEREST ME THE MOST IS REALLY TRYING TO UNDERSTAND THE ROLE OF ARCHITECTURE AND HOW CELL AGGREGATES RESPOND. IN LOOKING INTO THE ARCHITECTURE -- -- THIS IS A BOVINE OVARY. THERE IS A HUGE RANGE OF SIZES OF PORES AND HERE YOU CAN SEE THAT IN THE CORTEX IT'S VERY SMALL PORES AND STIFFNESS IS HIGHER AND WE BELIEVE THAT ALSO THAT LEADS TO MAINTAINING -- AND THERE IS A TRIGGER A REMODELING HAPPENS AND MIGRATION AND THESE ARE REMNANTS. WHERE THE MOST MATURE FOLLICLES RESIDED. BEFORE OVULATION. SO THEY ARE AT THEIR BIGGEST. SO THEY GO FROM VERY SMALL IN THE ORDERS OF 10 OF MICRONS ALL THE WAY TO MULTI-CENTIMETERS. SO THAT ACTUALLY ALSO WE HAVE TO THINK ABOUT THE DESIGN OF HOW CAN WE CREATE STRUCTURES THAT CAN ACTUALLY ACCOMMODATE THE HUGE GROWTH FROM PRIMORDIALS TO MATURE FOLLICLES. SO THAT * LED TO THE FIRST STUDIES WHICH WAS FOCUSING ON ARCHITECTURE. WE HAD TO START SOMEWHERE. WE LOOKED AT THE NATURAL OVARY. THE FOR SIZE AND THE ACTUAL ACCOMMODATION OF GROWTH AND WE WANTED TO SEE HOW DOES ARCHITECTURE AFFECT THE FOLLICLE VIABILITY AND FUNCTION. AND THE SIMPLE EXPERIMENT WE COMPARED DIFFERENT ADVANCING ANGLES. THIS IS A RELATIVE ANGLE BETWEEN ADJACENT PARALLEL STRUTS SO 30, 60 AND 90-DEGREE ANGLES WERE COMPARED. SECONDARY MASS FOLLICLES AND THERE WAS A VERY BIG DIFFERENCE IN THE SURVIVAL OF THESE FOLLICLES DEPENDING ON THE ARCHITECTURE. SO WHAT WE SAW IN THE ACUTE ANGLE STRUCTURES THAT THE FOLLICLES TEND TO WEDGE ITSELF IN THE CORNERS WHEREAS IN THE 90-DEGREE ON ES THEY TENDED TO STAY IN THE STRUTS AND THE IMPLICATION IS THE ONES THAT IS WERE WEDGED IN THE CORNERS ACTUALLY SURVIVED BETTER MAINTAIN THE SPHERICAL NATURE. SO -- PERCENT OF SURVIVAL OF THE FOLLICLES -- WAS OBVIOUS IN THAT YOU DO NEED MULTIPLE CONTACTS IN ORDER TO MAINTAIN THE SURVIVAL OF THE FOLLICLES INVITRO AS WELL AS SHOWING THAT THE ARCHITECTURE SO THE ONES -- THAT WERE MORE TORTUROUS -- THAT LED TO GREATER NUMBER OF CONTACTS WITH THE FOLLICLE ITSELF LED TO HIGHER VIABILITY. SO WE WERE ABLE TO CULTURE THESE FOLLICLES INVITRO AND THESE DIFFERENT ENVIRONMENTS AND THEY WERE ABLE TO GO FROM THE SECONDARY ALL THE WAY TO PRE-- STAGES. WE SWITCHED. THIS IS 6-8 DAYS OF FOLLICLE CULTURE WHERE WE SAW THEY WERE ABLE TO GROW UP TO APPROXIMATELY 70 MICRONS IN DIAMETER AND THEY RELATED IN FULLY M2 -- MATURE EGGS THAT WERE READY FORGETTER SITUATION AND THIS SHOWS THE INDICATION WHERE YOU SEE -- FROM THE FOLLICLE AND IF YOU LOOK AT THE CHROME 10 -- * THIS IS AN INDICATION OF M2 EGG AND SPINDLE LIKE GRAIN STRUCTURES HERE. WHY IT WORKED I THINK THIS IS A STRUCTURE THAT WAS MADE FROM GELATIN. A SOFTER MATERIAL BUT IT CAN BE REMODELED. 3-D PRINTED AND THIS NOT ONLY CREATED AN ARCHITECTURE TO ALLOW FOR A SUFFICIENT NUMBER OF CONTEXT BUT ALLOWED FOR THE GROWTH. SO THE SPACE AND THE FOR SIZE ALLOWED FOR THE GROWTH THAT IS NECESSARY TO GO THROUGH THE PROCESS OF MATURATION. AND THE ABILITY OF THE MATERIAL TO ACTUALLY GIVE THE RIGHT COMPLIANCE SO IT COULDN'T BE TOO STIFF THAT INHIBITED THE GROWTH BUT ALSO CAN BE REMODELED OVER TIME TO ACCOMMODATE THE LARGER FOLLICLES IS VERY IMPORTANT. SO WHAT WE DID WE WERE ABLE TO SHOW THE VIABILITY AND USE A SPECIFIC ARCHITECTURE THAT WORKED INVITRO. BUT WE WANTED TO SEE IF WE IMPLANTED THESE FOLLICLES. WE TOOK A MOUSE AND TOOK OUT THE OVARIES. AND THEN WE TOOK GREEN FLUORESCENT PROTEIN -- FOLLICLES FROM A MOUSE AND WE THEN SEEDED IT INTO THE 60-DEGREE ARCHITECTURE AND IMPLANTED THAT INTO THE STERILIZED MOUSE AND THIS IS WHAT WE CALL A BIOPROSTHETIC OVARY AND OVER TIME THIS IS ANNEX PLANT AND -- WE TOOK IT AND SAW THAT THERE ARE SOME PRY MORPHEDDIAL FOLLICLES BUT * WE DID SEE FOLLICLES AT DIFFERENT STAGES. EVEN THE ONES THAT GO UP TO THE -- FOLLICLE STAGE. WE ALSO SAW A GREAT AMOUNT OF VASCULARIZATION WITHIN THE TISSUE WITHIN THE IMPLANT. AND ALSO SYSTEMATIC RELEASE OF HORMONES SHOWED THAT THE ONES THAT HAVE RECOGNIZED MICE THAT DIDN'T HAVE THE IMPLANT SHOWED NO HORMONE LEVELS BUT THE ONES WITH THE IMPLANT THE HORMONE LEVELS WERE PROPERLY RECOVERED. WE TOOK SOME OF THE MICE AND MATE IT AND WHAT WE SAW IS THAT THE -- THIS LED TO ACTUALLY THE BIRTH OF THE FIRST BIOPROSTHESIS DE-- DERIVED PUP AND THIS RESIDUAL IMPLANT STILL WAS FUNCTIONING BECAUSE IT WAS PRODUCING THE NECESSARY HORMONES TO BE ABLE TO INDUCE LACTATION OF THE MOM SO THE PUPS WERE RAISED BY THE MOM UNTIL WEANING. AND YOU CAN SEE THERE IS MILK IN THE BELLY THERE. AND SO THE DEVICE ITSELF THE IMPLANT WAS STILL HORMONALLY FUNCTIONAL AT BIRTH. SO THE PUP THAT WAS DERIVED FROM THE GREEN FLUORESCENT PARTICLES MADE IT AND THAT GENERATED THE SECOND GENERATION. THESE PUPS -- THIS IS AN INDICATION THAT THIS PUP THIS INITIAL PUP GREW TO ADULTHOOD WAS MATE AND IT DEMONSTRATED THAT IT HAD NORMAL FERTILITY. THIS IS REALLY EXCITING BECAUSE ONE, IT WAS USUALLY IN THIS TYPE OF WORK THE FIRST TIME IT DOESN'T WORK AND THERE ARE MULTIPLE ITERATIONS. BUT IT ACTUALLY DID WORK. THIS VERSION 1.0 DID WORK SO THERE IS EXCITEMENT TO BE ABLE TO USE BIOPRINTING TO BE ABLE TO DESIGN DIFFERENT SCAFFOLDING IN ORDER TO CREATE A DESIRED BIOLOGICAL RESPONSE. TO GIVE YOU AN IDEA OF WHAT HAD TO HAPPEN IN ORDER TO WORK IT IS REALLY A MULTITUDE OF DIFFERENT THINGS FROM SEEING THE FOLLICLES WITHIN THE STRUCTURE, THE FOLLICLES HAD TO SURVIVE. THEY HAD TO GROW AND FUNCTION REGULARLY AND UPON IMPLANTATION THEY HAD TO SYSTEMATICALLY SECRETE SPECIFIC HORMONES THAT ALLOWED THE EGG TO OVULATE. IT WAS IMPLANTED AVENUE HAD TO TRAVEL THROUGH THE AT TH -- AT THE LOANIAN TUBE * SO THERE IS A LOT OF DIFFERENT EVENTS THAT HAD TO OCCUR IN ORDER FOR THIS TO WORK. *. THIS IS OUR VERSION 1.0 AND NOW WE'RE MOVING TOWARD MORE SOPHISTICATED STRUCTURES THAT CANNOT ONLY SUPPORT THE GROWTH AND VIABILITY BUT HOW CAN WE CONTROL WHAT FOLLICLES GET -- WHICH ONES MATURE AND THAT IS TO CREATE DIFFERENT MICRO ENVIRONMENTS. AS I MENTIONED WE UNDERSTAND WHAT IS IN THE NATURAL. OVARY AS FAR AS COMPARTMENTALLATION. * SO OUR GOAL IS LONGEVITY. CAN WE HAVE ARTIFICIAL OVARY UNDERGO NORMAL CYCLING. WE LUCKILY HAVE A PLATFORM -- THAT CAN REALLY BASKETBALL TUNEABLE TO CREATE DIFFERENT STIFFNESS. THESE ARE JUST SOME OF THE I THINK ONE OF THE THINGS THAT WE FOCUSED ON EARLY ON IS REALLY TRYING TO CREATE VERSATILE MATERIALS PALLET. A SYSTEM THAT WE CAN USE IN ORDER TO CREATE AND TUNE AND OPTIMIZE THESE MICRO ENVIRONMENTS. SO WE'VE BEEN ABLE TO DO A LOT SENSE THEN. AND WE ARE TRYING TO KEEP IN MIND THAT THE TRANSLATIONAL ASPECTS. TRYING TO UNDERSTAND HOW DOES THIS INITIAL -- DATA ACTUALLY HOW CAN WE THEN CREATE PRODUCTS THAT CAN MAKE A DIFFERENCE IN THE FERTILITY SPACE NOT JUST THE HOLY GRAIL OF COMPLETELY ARTIFICIAL OVARY THAT CAN BE IMPLANTED BUT ALSO OTHER PRODUCTS THAT ARE MORE -- LIKE CREATING MICRO ENVIRONMENTS FOR -- OR HORMONE REPLACEMENT. THERE ARE DIFFERENT AVENUES THAT WE'RE EXPLORING. AND ALSO KEEPING IN MIND THE MANUFACTURERBILITY AND THE REGULATORY PATHWAYS THAT IS NECESSARY IN ORDER TO BRING PRODUCTS TO CLINIC. THANK YOU FOR YOUR ATTENTION AND I LOOK FORWARD TO THE PANEL DISCUSSION. >> THANK YOU SO MUCH. SO I THINK WE WILL MOVE TO THE NEXT SPEAKER. NEXT SPEAKER IS DR. ABRAHAM. -- HE IS AN ASSOCIATE PROFESSOR AT PENN STATE UNIVERSITY. HIS MAJOR AREA OF RESEARCH IS IN THE FIELD OF 3-D PRINTING. HE HAS BEEN WORKING ON SEVERAL ASPECTS OF BIOPRINTING SUCH AS THE PROCESSES BY -- PRINTERS AND TISSUE. MATURATION FOR MANUFACTURING OF MORE THAN A DOZEN TISSUES AND ORGANS. DR. -- IS A LEADING SCIENTISTS WITH OVER 150 PUBLICATIONS INCLUDING AN AUTHOR. HE HAS RECEIVED SEVERAL INTERNATIONAL AWARDS. THE TITLE OF DR. -- TALK IS CHALLENGING AND PROMISE OF BIOPRINTING AND TISSUE RECONSTRUCTION. >> THANK YOU SO MUCH FOR THE GREAT INTRODUCTION AND THE INVITATION. SO LET ME SHARE MY SCREEN. SO AS REQUESTED I WOULD LIKE TO FOCUS ON THE CHALLENGES AND PROMISE OF BIOPRINTING FOR TISSUE RECONSTRUCTION. SINCE THIS IS A BROAD TOPIC. I'VE BEEN IN THE FIELD FOR ABOUT 15 YEARS AND IN THE LAST 10 YEARS THERE HAS BEEN AMAZING PROGRESS AND IT'S TOUGH TO DISCUSS THIS IN 15 MINUTES SO I'LL JUST FOCUS ON TWO MAJOR APPROACHES. ONE IS AS DR. LEWIS EXPLAINED WITH THE BIOPRINTING OF CELL AGGREGATES -- AND THE OTHER ONE IS THE -- BIOPRINTING PROCESS SO I'M GOING TO FOCUS ON THESE TWO MEASURE AREAS. SO IN THE LAST 15 YEARS -- OUR FOCUS IS ON -- BIOPRINTING PROCESSES. BIOPRINTERS AS WELL AS APPLICATIONS FOR MORE THAN A DOZEN TYPE OF TISSUES. THESE ARE RANGING FROM MUSCULAR SKELETAL TISSUE -- AND WE HAVE APPLICATIONS LIKE PRINTING LUNG TISSUE. CARDIAC TISSUE MODELS. MODELING TUMOR MODELS. IN THE MEANTIME WE ALSO WORKING ON BIOPRINTING OF COMPOSITE TISSUES WHICH IS MADE OF MULTIPLE TYPES OF THESE TISSUE TYPES. SO I DON'T WANT TO GO INTO THIS AS EVERYBODY HERE -- PRETTY MUCH KNOWS ABOUT BIOPRINTING AND KNOWS WHAT BIOPRINTING IS BUT IN THE LAST FIVE OR SIX YEARS IT'S BEEN VERY POPULAR. IN THE PAST WE USED TO PRINT IN AIR WHICH MAKES THE PRINTING PROCESS CHALLENGING AS WE DEAL WITH SOFT MATERIALS AND THEY CAN EASILY COLLAPSE DOWN AND WE CANNOT MAKE THE STRUCTURE INTEGRITY. SO WITH THE BIOPRINTING WE CAN GENERATE STRUCTURES THAT ARE ANATOMIC AND COMPLEX. AND WE CAN GENERATE HIGH RESULT STRUCTURES USING BIOPRINTING. THERE ARE MULTIPLE TECHNIQUES. BUT IN GENERAL BIOPRINTING HAS QUITE MANY ADVANTAGES LIKE FOR EXAMPLE WE CAN CONTROL THE -- THE COMPOSITION DEPENDING ON THE TECHNIQUE -- WHERE WE CAN CREATE VERY SMALL TINY DROPLETS 30 OR 40 MICRON DROPLETS AND WE CAN CULTURE CELLS. MULTIPLE CELLS AND NOW WE HAVE THE CAPABILITY -- AS THE PREVIOUS -- DEMONSTRATED. AND WE HAVE THE ABILITY TO PRECISELY CONTROL THE SIZE AND WE CAN HAVE HIGH RESOLUTION. AND THEN ONE OF THE OTHER IMPORTANT FACTS AND PARTICULARLY -- FOR EVERYTHING FROM PHARMACEUTICAL AND DRUG TESTING IS THE ABILITY TO CREATE AND PRINT A NUMBER OF TISSUES IN -- CAN BE USED AND THAT CAN BE USED FOR DRUG TESTING. SO THERE ARE TWO MAJOR APPROACHES THAT WE HAVE IN BIOPRINT APPROXIMATING WHETHER WE USE A SCAFFOLD. WE DON'T REALLY USE A SCAFFOLD. THEY HAVE ADVANTAGES AND DISADVANTAGES WITH RESPECT TO EACH OTHER SO I PROVIDED TWO EXAMPLES. ONE FROM ADAM AND THE OTHER FROM -- SO IN THE FIRST ONE THE FIRST WE SEE -- A 3-D OF THE HEART IN HIGH RESOLUTION. VERY TINY FEATURES CAN BE CREATED IN THE -- THROUGH THE ENTIRE -- OF THE HEART CAN BE EASILY MIMICKED. THESE ARE THE BLOOD VESSELS THAT ARE PRINTED RANGING FROM 10 MICRONS TO TO-30 MICRONS. INSTEAD OF USING A SCAFFOLD -- DURING THE PRINTING PROCESS. THE CELL AGGREGATES CAN BE BIOPRINTED. -- TISSUE STRANDS WHICH IS SOMETHING THAT WE DEMONSTRATE IN THE PAST CAN BE USED TOO. SO WITH THIS CELL AGGREGATES THESE CAN BE PRINTED -- WITH THE HELP OF A MOLD STRUCTURE. AS YOU CAN SEE HERE THE -- THE PRINTED -- CONFUSE INTO A SINGLE PATCH OF TISSUES. THIS IS A NICE CROSS SECTION -- SO THIS CAN BE A FURTHER CULTURE. IT HAS SUFFICIENT INTEGRITY. THE GOOD THING -- AGAIN WE CAN GENERATE ANY COMPLEXITY THAT WE WANT. IT'S A SCALABLE PROCESS. WE CAN MAKE HUGE STRUCTURES. IT'S RAPID -- PROCESS. SO ANY PERSON WITH MINIMAL EXPERIENCE CAN USE THIS IN A SUPPORT PATTERN AND CAN MAKE COMPLICATED SHAPES BUT THE PROBLEM IS THE CELL DENSITY -- HAVE TO WHITE QUITE A LONG TIME TO FINISH. THE PROCESS. BUT YOU'RE USING HIGH CELL DENSITIES. AND YOU -- AND IT DOESN'T HAVE AGGREGATION. BUT THE APPROACH IS NOT EASILY SCALABLE. THERE IS LIMITED RESOLUTION AND NOT PRACTICAL. SO IN THE MEANTIME -- YOU HAVE NEED TO CULTURE QUITE A LOT OF CELLS AND DEDICATE ALL OF YOUR TIME CULTURING CELLS AND THIS TAKES A LOT OF TIME TOO. SO WE USE BOTH TECHNIQUES AND TODAY I'M GOING TO FOCUS ON ONE OF THE TECHNIQUES THAT WE INTRODUCED. SO WE CAN DO THAT BY PRINTING THESE SPEAR ROADS ON TOP OF EACH OTHER. WE CAN MAKE COMPLICATED STRUCTURES. THE SPHEROIDS DON'T HAVE TO BE THE SAME SIZE. WE CAN MAKE COMPLICATED STRUCTURES. AND USING THIS TECHNIQUE WE CREATED DIFFERENT TISSUE TYPES LIKE TISSUE MODELS FOR EXAMPLE -- THE INTERFACE WHERE WE HAVE A CLEAR INTERFACE. THIS WAS MADE USING THE STEM CELLS AND WE'RE DOING THE SAME WITH THE STEM CELLS BUT NOT CULTURING THEM IN A DIFFERENTIATED MEDIAN. DIFFERENTIATE THE CELLS AFTER PRINTING INSTEAD OF DIFFERENTIATING THEM BEFORE PRINTING. SO, IT'S NOT -- THE METHOD OF PRINTING OF SPHEROIDS BUT WE CAN ALSO PRINT STRANDS. THESE STRANDS ARE A DIFFERENT FORM OF THE SPECIAL RIDE. WE SAW THAT THE ORIENTATION IN THESE STRANDS ARE LONGITUDINAL. THEN WE ACTUALLY 3-D PRINTED THESE VERTICALLY AND HORIZONTALLY AND OVER TIME THEY ASSEMBLED INTO A SINGLE PATCH OF TISSUE AND OUR GOAL WAS TO MIMIC IT WHERE WE HAVE THE FIBERS -- INVERTED VERTICALLY IN THE DEEP ZONE OF THE CARTILAGE VERSUS THE COLLAGEN ORIENTATION THAT IS PARALLEL TO THE TOP SURFACE IN THE SUPERFICIAL ZONE. SO WE FURTHER ADVANCED THIS AND DEMONSTRATED THAT WE CAN USE THIS TECHNIQUE IN A FREE FORM MATTER. SO WE CREATED THE DEVICE WHERE WE HAD THESE SPEAR RIDE CONTAINERS. 7 AND WE PLACED THEM INTO THE GEL WHICH HOLDS THE SHAPE AND LET THE SPHEROIDS FUSE AND MAKE A 3-D STRUCTURE. SO USING THIS WE CAN MAKE COMPLICATED SHAPES THAT HAS BEEN DEMONSTRATED LIKE MAKING THESE KIND OF SPIRAL -- TYPES OF PATTERNS. SO WE ALSO USED THAT FOR MORE BIOLOGICAL EXAMPLES AND DIFFERENT GELS. WE USE -- OTHER SUPPORT GELS -- WHERE WE PRINTED THE -- --th BONE SPHEROIDS. THESE TWO WERE DIFFERENTIATED USING THE TECHNOLOGY SO WE SPECIFICALLY USED -- FOR THE 3-D PRINTING OF THE BONE AND -- PRINTING OFF THE -- SPHEROIDS. SO WE MADE THESE STRUCTURES MIMICKING. ONE OF THE PROBLEMS WITH THIS -- TECHNIQUE THAT WE TRIED TO -- WE SPENT A LOT OF TIME TRYING TO SOLVE THIS -- -- [ INDISCERNIBLE ] THESE SPHEROIDS OF COURSE THEY PRODUCE A SIGNIFICANT AMOUNT OF COLLAGEN FIBERS. AND THEN OVER TIME IF YOU DON'T CONTROL THE PROCESS CAREFULLY THEN YOU PRINT A RING FOR EXAMPLE THESE ARE THE CARTILAGE SPHEROIDS AND IT TURNS INTO A BALL SHAPE AND YOU LOSE YOUR FIDELITY. SO ON THE OTHER HAND YOU CAN USE OTHER APPROACHES LIKE YOU CAN BLOCK THE COLLAGEN OR YOU CAN CONTROL THE DIFFERENTIATION PROCESS SO THAT THE CELLS CAN BASICALLY MOVE INTO THE DIFFERENTIATION PATHWAY. AND PROLIFERATING. AND THEN THE COMPASSION. COME -- COME PACKS. YOU ARE LIMITED TO THE SPHEROID SIZE. AS YOU MAKE SMALLER SPHEROIDS YOUR RESOLUTION CAN BE BETTER AND FOR SCALABILITY -- THE PRINTING CAPABILITY IS WHERE WE CAN PRINT SAY 150 SPHEROIDS AT A TIME. INSTEAD OF MAKING THEM ONE BY ONE. WE FURTHER INTEGRATE THIS WITH THE SCAFFOLD BASED SYSTEM AS WELL. WHAT YOU SEE IS INTEGRATION -- OF THESE SYSTEMS TOGETHER. FOR MICRO PHYSIOLOGICAL SYSTEMS. SO THE PRINTING CAN ENABLE THE DEPOSITION OF THE -- SPHEROIDS. AND AS YOU CAN SEE HERE YOU CAN ALSO GET THOSE -- BY CONTROLLING THE PRINTING PARAMETERS LIKE THE SPACE AND THE SUPPORT CELLS ARE ON THESE PRINTED SPHEROIDS SO WE PRINTED THEM INTO A DEVICE WHERE WE HAVE A CHANNEL AND AS YOU CAN SEE HERE THE TUMOR -- [ INDISCERNIBLE ] AND THESE -- THE MAIN CHANNEL AND WE CAN SEE THAT THE CANCER CELLS ARE INVADING INTO THE CAPILLARIES AND THEY ALSO METASTASIZE THROUGH THIS CHANNEL. SO WE HAVE TWO OF STRATEGIES. ONE IS TESTING THE -- -- -- AND THEN AS YOU CAN SEE HERE BY APPLYING DIFFERENT CONCENTRATION OF THE DRUG WE CAN DEFINITELY CONTROL THE TUMOR KILLING PROCESS. WE'RE ALSO USING T CELLS AND PROFUSING T CELLS AND EFFECTIVELY KILLING THESE TUMORS WITH THE -- WE COLLABORATE WITH I'M NOT JUST -- AND WE'LL HOPEFULLY HAVE THIS SOON. -- -- INTO THE DEFECT INTO A SURGICAL SETTING. THIS WAS FIRST ORIGINALLY PROPOSED BY LEE WISE WHEN HE WAS -- BACK IN 2007 SO AT THAT TIME THIS CONCEPT PRINTING THE GROWTH FACTORS AND THEY HAD A PILOT STUDY SHOWING JUST THE PRINTING ON AN ANIMAL. AND THEN LATER THIS WAS -- APPLIED BY DIFFERENT GROUPS INCLUDING -- IN PRINTING THE SKIN TISSUE IN MULTIPLE LAYERS. AND 3-D SCANNING IN SMALL AND LARGE ANIMALS AS WELL AS USING THE LASER ASSISTED BIOPRINTING BY THE GROUPING -- SO IN FRANCE THEY USED THE LASER SYSTEM TO DEPOSIT THE DROPLETS ON THE -- OF MICE. AND THAT WAS -- A STUDY PUBLISHED SHOWING THE -- DEPOSITION. INSTEAD OF USING A BOT A HAND-HELD SCANNER WAS USED. -- THE INJECTED GEL CAN BE WITH THE CELLS AND WITH THE -- LIGHT THE POLARIZATION CAN BE CONTROLLED AND DIFFERENT SHAPES OF TISSUES CAN BE PRINTED. SO WHAT WE DID WE FURTHER ADVANCED THESE TECHNOLOGIES SO WE FOCUS ON TISSUE -- WE HAD TWO DIFFERENT TYPES. ONE IS THE BONE AND THE OTHER IS SKIN. FOR THE BONE TISSUE WE CREATED A BONE INK -- AND FOR SOFT TISSUE WE USED -- COLLAGEN AND THEN WE USED DIFFERENT BIOLOGY LIKE -- AND WE HAVE OTHER PUBLICATIONS GOING ON WITH THE GEN -- GENE THERAPY. THAT DEFECT GOES DOWN TO THE DURA. THE DURA ON THE BRAIN WAS KEPT INTACT AND ON TOP OF THAT WE HAD A BURIAL LAYER THAT PREVENTS THE SAID MEN TAKES OF THE CELLS IN THE SOFT TISSUE LAYER. -- SO WE HAD THE 3-D SCANNER AND SCANNING INFORMATION IS BEING SENT TO THE BIOPRINTER AND THE BIOPRINTER IS PRINTING THE BONE TISSUE FIRST. SO FIRST WE TESTED INDIVIDUAL TISSUES. SO WE TESTED BOTH THE BONE PRINTING -- AS YOU CAN SEE HERE THESE ARE THE RESULTS FOR SIX WEEKS. SO WE HAVE ABOUT 75% TO 80% AREA COVER AM. WE DIDN'T USE ANY CELLS. -- -- AND THEN WE ALSO LOOK AT THE SKIN REGENERATION. SO VARIOUS DIFFERENT GROUPS BUT THE GROUPS THAT WE HAVE WITH THE -- ---IN THE DERMAL LAYER -- -- -- YIELD THE BEST RESULTS IN TERMS OF THE HEALING RATE IN TERMS OF THE REACTION VASE POTENTIAL AND WE LOOK AT THE MECHANICS. WE COMBINED AND CREATED DEFECT ALL THE WAY DOWN TO THE BRAIN AND THE DURA WAS INTACT. AND WE PRINTED THE OTHER THREE LAYERS. AND AS YOU CAN SEE HERE THE WOUND HEALED IN ABOUT FOUR WEEKS. WE DIDN'T GET -- PROBLEMS WITH -- BECAUSE THE SKIN DEFECT WAS RELATIVELY SMALL. OF COURSE VERY LARGE SKIN DEFECTS CAN BE USED BUT WE WANTED TO SEE HOW THE INTERFACE WAS FORMING. SO WE LOOKED AT THE BONE REGENERATION. AT WEEK SIX WE GOT BETTER BONE REGENERATION. WHICH WAS A DAY LESS BUT THE BONE REGENERATION WAS LESS COMPARED TO THE BONE ONLY -- BECAUSE IT'S OPEN TO AIR. -- WE ACTUALLY -- SUTURED THE SKIN ON TOP AND IT PROVIDES A LOT OF VASCULARIZATION. WE HAVE A LARGE PROJECT THAT WE'RE GOING DEEPER AND TRYING TO UNDERSTAND HOW WE SHOULD -- ---AND ADVANCE THE PROCESS. LIMITATIONS. BIOPRINTING -- IS REALLY CHALLENGING. IT'S NOT JUST PRINTING -- AND OF COURSE PRINTING MATERIALS IS PRETTY STRAIGHTFORWARD BUT IF YOU WANT TO GET FUNCTIONAL OUTCOMES YOU WANT TO COMPARE THE COMPOSITE TISSUE RAPIDLY AND THAT IS REALLY IMPORTANT. IN THE MEANTIME WE ARE ---WE ALSO NEED VASCULARIZATION. WE COULD BE ABLE TO ADVANCE THE PROCESS. SO NORMALLY IT TAKES ABOUT TWO WEEKS SO -- APPROACH THAT WE DEMONSTRATED WITH THE MICRO PUNCTURE. A NEW SURGICAL TECHNIQUE THAT WE CREATED WITH COLLABORATION FROM THE SURGEONS IN -- HERSHEY AT PENN STATE. 36-48 HOURS IN JUST TWO DAYS. THIS ACTUALLY HELPS US WITH -- BIOPRINTING BUT FOR VERY LARGE ORGANS WE NEED TO CONTROL THE VASCULARIZATION. AND EMBED THE EXISTING BLOOD VESSELS IN THE PROCESS. SO THE OTHER INTERESTING THING IS OF COURSE -- BIOPRINTING TISSUES ON THE NONLOAD BEARING SIDES IS EASIER THAN PRINTING FOR THE LOAD BEARING SIDES. FOR BONES YOU ALSO NEED MECHANICAL SUPPORT. YOU CANNOT SIMPLY PRINT SOMETHING THAT WILL FILL THERE BUT NATIONALS YOU NEED SOMETHING THAT IS STRUCTURAL AND STABLE AND THAT IS ALSO CHALLENGING. FOR DEEP ORGANS THERE IS A NEW INTEREST -- AND PUBLISHED. IT REQUIRES A MINIMUM INVASIVE CASE. WHERE A LITTLE INCISION IS CREATED HERE AND THEY USE -- -- WHICH IS -- SO YOU CAN CONTROL THIS WITH MAGNETS AND THEY COULD BE BE ABLE TO PRINT BY CONTROLLING THE POSITION OF THIS WITH THE MAGNETS. THE NOZZLE TIP. THE PRINTING PROCESS WAS CONTROLLED. IT WAS VERY CHALLENGING SO THIS IS UNDER THE SKIN ONLY BUT FOR VERY DEEP ORGANS THAT ARE NOT FLAT IN SURFACE. THIS IS STILL A VERY CHALLENGING TASK. SO WITH THAT -- I ALSO WOULD LIKE TO EMPHASIZE THE PROMISES OF THIS -- PRINTING SO WE WENT THROUGH THE LIMITATIONS. SO NOW WE'RE WORKING ON LARGE ANIMALS. FOR LARGE ANIMALS WE'RE NOT USING THREE DIMENSIONAL -- BUT RATHER -- A ROBOTIC ARM IN COLLABORATION. SO, THAT IS GOING TO BE FOR REPAIRING THE CRANIAL DEFECTS ON SHEEP. AND OF COURSE THE GOAL IS TO TRANSLATE THIS AND ONE DAY SEE BIOPRINTERS IN CLINICS. LIKE THIS WAS THE COVER OF MY BOOK THAT SHOWS PRINTING MULTIPLE DIFFERENT TISSUE TIMES IN A CLINICAL SETTING. SO IF THIS WILL WORK -- -- THIS IS BACK IN 2015. WHERE I WAS UNDER A 3-D PRINTER IN THIS POSITION. I WAS REQUESTED TO SEE HOW I FEEL. OF COURSE. THIS WAS A 3-D PRINTER THAT WAS PRINTING -- AND IT WAS A LITTLE SCARY EXPERIENCE. OF COURSE IN THE CLINICS WE DON'T EXPECT THIS KIND -- THE MAXIE -- -- ---BUT I THINK CURRENT SURGICAL -- CAN BE EQUIPPED WITH BIOPRINTING TIPS AND THEY CAN EASILY PRINT THINGS ON HUMANS IN THE FUTURE. WITH THAT I WOULD LIKE TO THANK EVERYBODY FOR THE ATTENTION AND -- THE MEMBERS AND COLLABORATORS WHO CONTRIBUTED TO THIS RESEARCH WHICH WAS FUNDED BY VARIOUS INSTITUTIONS AT NIH. >> THANK YOU. SO THANK YOU FOR THE VERY RICH PRESENTATIONS. SO BEFORE WE OPEN FOR THE PANEL DISCUSSION -- I WOULD LIKE TO CHECK WITH -- . CAN WE RUN OVER TIME HERE? >> WE HAVE SOME BUFFER TIME. IN THE BREAK. SO WE CAN USE THE BREAK -- MAYBE COMPRESS THE BREAK TIME SO YOU CAN -- I THINK WE ENCOURAGE DISCUSSION. SO ABSOLUTELY. WE HAVE A DEADLINE. >> OKAY. SO FIRST OF ALL THERE IS A QUESTION -- FROM -- AND SO TO DR. LEWIS. DR. LOUIS HAS ADDRESSED BY THE CHAT BUZZ FOR THE BENEFIT OF THE AUDIENCE WHO JOINS ON THE VIDEO CASTLED LIKE DR. LEWIS TO ADDRESS THIS QUESTION. AND THEN PLEASE FEEL FREE TO JUMP IN AT ANY TIME. THE QUESTION IS CAN YOU EXPAND YOUR THOUGHTS ABOUT THE SACRIFICE COLIN CANS AND WHY IT'S IMPORTANT TO BIOPRINT CELL RATHER THAN THE SCAFFOLD >> THE LAST TALK TALKED ABOUT THIS. IT DOES DEPEND ON THE TISSUE TYPE. AND WHAT YOU'RE TRYING TO -- REPAIR OR REGENERATE. SOME TISSUES LEND THEMSELVES NATURALLY TO SCAFFOLD BASED APPROACHES AND OTHERS MAYBE NOT SO. SO WE'VE BEEN FOCUSING ON COMPLEX ORGANS LIKE THE HEART AND THE KIDNEY AND I WOULD ARGUE IN THOSE CASES IT'S DIFFICULT TO START WITH A D CELL RECELL APPROACH OR FRESH APPROACH TO PRINTING A SCAFFOLD AND THEN SEEDING IT WITH CELLS. SIMPLY BECAUSE THE ARCHITECTURE OR COMPLEXITY THAT IS NEEDED IS SO SOPHISTICATED. BUT IN OTHER EXAMPLES THAT IS PROBABLY NOT THE CASE. >> THANK YOU. SO I HAVE A QUESTION. AND -- WITH NICHD -- I REALLY LIKE THE SPEAKER TO TELL US -- WHEN THE FIELD EXPANDS INTO THE PEDIATRIC POPULATION EVEN TO THE BABIES. WHAT -- WILL BE DONE. THE CHALLENGING AND ALSO THE ADVANTAGE AND DISADVANTAGE AND ASSOCIATE CHALLENGES. >> IF YOU DON'T MIND -- JUST TAKE THE FIRST SWING AT THIS AND I WOULD SAY STARTING WITH ORGANOIDS AND THESE STEM BASED PROTOCOLS YOU GET QUITE YOUNG TISSUE. SO, THAT IS A LIMITATION OF THAT APPROACH. IF YOU'RE NOT USING A MULTI-CELLULAR AGGREGATE THAT IS MATURE TO BEGIN WITH. BUT IT'S A POTENTIALLY A VERY POWERFUL METHOD FOR ONE CHILDREN OR BABY TYPE DEFECTS. SITES A REALLY GOOD PLACE TO START FOR A SWIFT BASED PRINTED TISSUE AND MAYBE OTHERS WANT TO CHIME IN. >> I THINK FOR PEDIATRIC APPLICATION IS SOMETHING THAT WE'RE ALSO LOOKING INTO. BUT ON THE -- ORGAN BUT THE MUSCULAR SKELETAL WHERE THERE IS FASTER APPLICATION LIKE BONE OR TRACHEA THAT HAS BEEN EXPERIMENT EXPERIMENTED ON BABIES AND ALSO ADOLESCENTS AND THAT IS WHERE WE PROBABLY START SEEING FIRST USES OF SOMETHING THAT IS 3-D PRINTED OR PATIENT MATCHED AND AND THE ABILITY OF THESE TISSUES TO ACTUALLY GROW WITH THE PATIENT WHICH IS ONE OF THE BIGGEST ADVANTAGES. SO THERE IS A REAL NEED IN THE PEDIATRIC SPACE TO REALLY SHOWCASE THAT 3-D PRINTED TECHNOLOGIES CAN BE USED FOR THESE PATIENTS AND THEY ARE SAFE AND GROW AND THEN MOVING ON TO MORE SOPHISTICATED TISSUES AND ORGANS. >> SO WE ACTUALLY HAVE SOME INTERESTING -- COULD THIS -- THAT IS THE DEFECT ON THE KIDS WHEN THEY ARE BORN AND IT DOESN'T HEEL -- NATURALLY SO THERE NEEDS TO BE TREATMENT. AND THE WAY THAT WE PROPOSE LIKE HAVING THE BONE PRINTED AND MULTIPLE LAYERS OF SKIN COULD EASILY REPAIR THE DEFECT. >> SINCE ALL OF -- THREE OF YOU ARE WORKING ON MULTIPLE TISSUES AND ORGANS AND SO I WOULD LIKE TO LEARN IN YOUR VIEW WHICH FIELD DO YOU THINK WE'RE CLOSER AND IN WHICH FIELD -- THERE IS MORE THAT NEEDS TO BE DONE. >> AND ALSO HOW CAN WE LEARN FROM EACH OTHER. >> WE'RE FARTHER AWAY THAN OTHERS AND A VERY GOOD POINT. SOME OF THE MUSCULAR SKELETAL AND TRACHEA, THOSE ARE NOT ONLY IN SOME CASES WE ALREADY HAVE 3-D PRINTED SOLUTIONS AND THEY'VE BEEN I'M PLANT AND SHOWN EFFICACY. SO IT SPANS A BROAD RANGE. >> I THINK THE MAJOR CHALLENGES THAT I'M EXPERIENCING NOWS HOW DO WE ACTUALLY CREATE REPRODUCIBLE PRODUCTS GETTING THROUGH THE REGULATORY APPROVAL FOR NEW PRODUCTS THAT ARE 3-D PRINTED WHEN IT COMES TO TISSUE ENGINEERED MATERIALS AND UNDERSTANDING THAT LANDSCAPE AND HOW IT FITS WITHIN THE OVER-ALL SYSTEM OF HEALTH CARE AND QUALITY AND REGULATIONS THAT ALSO HAVE TO BE SQUARED AWAY BEFORE IT IS USED MORE PREDOMINATELY IN PATIENTS SO NAVIGATING WHAT THAT LOOKS LIKE AND UNDERSTANDING WHAT THAT LOOKS LIKE IN THE VERY BEGINNING OF DESIGN AND R & D. IT'S HUGE TO BE ABLE ABLE TO KNOW THAT IN ORDER TO BE PREPARED. NO MATTER WHAT STRUCTURE YOU'RE TARGETING. >> SO WE SEE MORE PRODUCTS AND WE WILL START SEEING THE PRODUCTS WITH THE -- ---AS WELL AS DRUG SCREENING. THERE IS THE REGULATIONS THAT ARE NOT THAT TIGHT AS THE PRINTED TISSUES TO BE TRANSPLANT INTO HUMANS. SO WITH THE DISEASE I THINK THERE IS A HUGE POTENTIAL PARTICULARLY UNDERSTANDING HOW THEY SAY CANCER GROWS OR OTHER TISSUES LIKE LUNG TISSUE THERE IS -- AND CREATING MODELS AT THE LOWER LEVEL AND SEE HOW THEY INTERACT WITH THE VIRUSES AND USE THEM AS A PLATFORM TECHNOLOGY TO UNDERSTAND HOW THE TISSUE INTERACTS WITH THE MICROBIOME AND THE VIRUS AND BACTERIAL INFECTION. THESE WILL TRANSLATE INTO MARKET SOONER THAN BIOPRINTED TISSUES. EVEN THE MORE COMPLICATED ORGANS HEART, AND LUNGS STILL I CAN SAY A COUPLE OF MORE -- BUT THERE ARE ORGANS LIKE THE BRAIN WHICH IS NOT FULLY UNDERSTOOD YET AND THIS WILL TAKE I BELIEVE A LONGER TIME. >> THANK YOU. I FEEL LIKE -- -- BECAUSE I WOULD LIKE TO KNOW WHAT WOULD BE THE LOW HANGING FRUIT THAT WE CAN ACHIEVE IN THE NEAR FUTURE. JUST A HUGE EFFORT. A DECADE TO REACH WHERE WE ARE NOW. BUT STILL -- I WOULD LIKE TO KNOW WHAT WE CAN ACHIEVE IN THE SHORT TIME AND ALSO WHAT ARE THE ROLE APPLICATION THAT YOU VISION IN YOUR RESPECTIVE FIELD. >> PROBABLY NOT THE BEST PERSON TO START WITH BECAUSE WE FOCUS OUR LAB ON THE HARDER CHALLENGES. AND WE DO HAVE SOME EFFORTS IN SKIN PRINTING AND THINGS LIKE THAT WHICH IS MUCH MORE NEAR TERM. BUT YEAH. I WOULD SAY THAT THE TWO DIMENSIONAL. THE SIMPLE SORT OF TUBULAR CONSTRUCTS LIKE A TRACKIAL SUPPORT * DEVICE. THOSE ARE EITHER ALREADY IN THE CLINIC OR ON THE NEAR HORIZON IN PRINTED FORMS. I THINK WHEN WE ARE CIRCLING AWAY FROM SIMPLE GEOMETRIES OR TWO DIMENSIONAL SHEETS THAT IS WHEN WE HAVE TO FACE A NUMBER OF OTHER CHALLENGES. AND WHEN YOU ADD REALLY COMPLEX ARCHITECTURES AND MULTIPLE CELL TYPES AND MULTIPLE FUNCTIONS LIKE THE KIDNEY FOR EXAMPLE THEN IT IS QUITE A LARGE LIFT >> SO -- HOW I SEE IT -- RIGHT -- 3-D BIOPRINTING CANNOT ONLY BE USED FOR ENGINEERING TISSUES BUT ALSO FOR OTHER HEALTH CARE INDICATIONS. FOR EXAMPLE ONE OF THE THINGS THAT WE'VE BEEN WORKING ON IN THE FERTILITY SPACE IS MATURATION AND INVITRO FIRST AND HORMONE REPLACEMENT THERAPY USING MORE BIOPRINTING AND INPUT IN USING THE PATIENT'S OWN CELLS. SO THERE ARE I GUESS PRODUCTS THAT I THINK CAN COME OUT IN THE NEAR TERM THAT DON'T REQUIRE IMPLANTATION PER SAY AND REGENERATION OF TISSUE BUT CAN STILL BE APPLIED IN THE HEALTH CARE SPACE. DIAGNOSTICS, DRUG SCREENING, CANCER THERAPIES. I THINK YOU'LL SEE THOSE EARLIER BECAUSE OF THE LESS REGULATORY BURDEN. BUT IF SOMETHING CAN SHOW EFFICACY OR THE ABILITY TO PREDICT PATIENT OUTCOMES I THINK THOSE HAVE POTENTIAL TO BE ON THE MARKET FASTER. EVEN AND AGAIN AS WE MENTIONED -- AND WHAT JENNIFER WAS SAYING THE LESS SOPHISTICATED TISSUE TYPES THERE ARE BONE GRAPHS OUT THERE AND ONE THAT'S MAYBE WORK BETTER. ONES THAT CAN BE INTERFACED WITH OTHER HARDWARE IN THE ORTHOPEDIC REALM THAT CAN LEAD TO BETTER OUTCOMES OR LESS COMPLICATIONS. SO THERE IS MORE -- A LOT OF INDICATIONS THAT ARE SIMPLER THAN JUST AN ORGAN REPLACEMENT THAT I THINK WILL COME OUT IN THE NEAR TERM. EVEN LIKE PATCHES. MAYBE SOMETHING THAT CAN SUSTAIN PATIENTS LONGER ON THE WAITING LIST BEFORE THEY GET A TRANSPLANT. SO I THINK THERE IS DEFINITELY ROOM FOR OTHER BIOPRINTED SOLUTIONS TO BE USED NEAR TERM >> GOOD POINT. >> I AGREE WITH ALL OF THESE COMMENTS. SO I CAN SAY THE VASCULAR CONSTRUCTS -- COULD BE THE NEW HORIZON. IN THIS REGARD I THINK CARTILAGE HAS A LOT OF EXISTING TRANSLATION NOT WITH THE BIOPRINTING BUT WITH NONBIOPRINTED CONSTRUCTS FOR -- REPAIR, FOR ARTHRITIS PURPOSES. I DON'T WANT TO SIMPLIFY THESE ORGANS. NO ONE HAS DEMONSTRATED THE NEEDED ULTRA STRUCTURE OF THE CARTILAGE. AND THE SKIN IN ITSELF. LIKE WE'RE TRYING TO GROW SKIN AND IN THE MEANTIME WE WOULD LIKE TO HAVE THE HAIR FOLLICLES AND SWEAT GLANDS. THEY DON'T REALLY GROW STRAIGHT. SO IT DOESN'T REALLY LOOK LIKE 100% IS WHAT THE NATIVE TISSUE IS BUT FOR REGENERATIVE MEDICINE PURPOSES IT COULD BE LIFESAVING. SO OTHER THAN THAT AS I MENTIONED THE ORGAN MODELS AND TISSUE MODELS I THINK IT WILL BE A GREAT STEP AND AN EXAMPLE FOR THE BIOPRINTED TISSUES AND -- THE TRANSLATION OF THE OTHER COMPLEX BIOPRINTED TISSUES FOR TRANSPLANTATION PURPOSES IN THE FUTURE. AND PERSONALLY I WOULD LIKE TO SEE BIOPRINTERS IN CLINICS NOT JUST -- BIOPRINTING BUT LIKE A CLEAN ROOM -- HAVING THE FACILITIES THAT CAN DIRECTLY USE STEM CELLS FROM PATIENTS AND MAKE THE REPLACEMENT PARTS AND TRANSPLANT THEM BACK INTO PATIENTS >> AND BUILDING ON RAMILLE SAID -- FOR EXAMPLE FOR KIDNEY REGENERATION REPAIR OR RENAL REPLACEMENT THERAPY WOULD NOT BE WHOLE ORGAN TRANSPLANTATION. A STEP WOULD BE GIVING A PATIENT 10% IMPROVED KIDNEY FUNCTION TO GET THEM OFF OF STAGE FIVE DIALYSIS UP TO STAGE FOUR TO STAGE THREE. SO THAT THEY HAVE A BETTER QUALITY OF LIFE AND A LONGER TIME AND SORT OF BRIDGING THE GAP TO WAIT FOR A DONOR ORGAN THAT WOULD BE APPRECIATE. THAT WOULD BE AN INTERMEDIATE STEP AND THE SAME FOR A MYOCARDIAL INFARCTION. REPAIRING A SPECIFIC WEDGE OF TISSUE THAT MAY HAVE GOTTEN DAMAGED DURING A HEART ATTACK. YOU WOULD NOT BE REPLACING THE WHOLE HEART SO THERE ARE PATHWAYS. SHORTER TERM PATHS TO DEMONSTRATE THAT THE BIOPRINTED AND IN OUR CASE A HYBRID TISSUING BIOLOGICAL SELF-ASSEMBLY COUPLED WITH BIOMANUFACTURING COULD BE A POTENTIAL SOLUTION. >> ABSOLUTELY. >> TALK ABOUT THE BETA CELL REPLACEMENT APPROACHES. AND OF COURSE PANCREAS -- HAS BOTH -- FUNCTIONS AND NOW MOST OF THE RESEARCHERS THEY FOCUS ON THE ENDOCRINE FUNCTION AND HOW WE CAN GENERATE A REPLACEMENT -- THAT CAN PRODUCE INSULIN. THAT IS THE FIRST STEP AND THEN -- OTHER FUNCTIONS LATER ON. WILL MAKE IT CLOSE TO FULL FUNCTIONING ORGAN. >> SO I WANT TO SAY THAT YOUR INPUT AND YOUR FEEDBACK IS VERY IMPORTANT FOR US. BECAUSE WE ARE THINKING ABOUT HOW TO PUT OUT AN INITIATIVE FOR POTENTIAL FUNDING. I WOULD LIKE TO KNOW THE SHORT-TERM GOAL WE CAN ACHIEVE. AND ALSO THE LONG-TERM GOAL. FOR THE FUTURE TO PROMOTE THE FIELD. I CAN REALLY GO ON. THIS IS SO INTERESTING BUT I THINK IN LIGHT OF THE TIME WE PROBABLY NEED TO TAKE A BREAK. AND -- >> YES. WE'LL MOVE ON INTO THE NEXT SESSION AND TO RESPECT THE TIME OF OUR SPEAKERS >> WE THANK THE THREE SPEAKERS FOR THEIR SLEPT PRESENTATIONS AND YOUR WISDOM. THANK YOU. >> I'M SORRY I FEEL LIKE I'M THE PARTY BREAKER BUT WE NEED TO MOVE ON. OUR NEXT SESSION IS -- SESSION 5 AND WILL BE MODERATED BY DR. MARTHA LUDBERG FROM THE NATIONAL INSTITUTE -- NATIONAL HEART, LUNG AND BLOOD INSTITUTE. SHE WORKS IN THE DIVISION OF CARDIOVASCULAR SCIENCE. MARTHA, TAKE IT AWAY. >> I'M EXCITED TO WELCOME OUR FINAL SESSION OF THE WORKSHOP FOR REAGAIN MEDICINE. PART B. THIS SESSION WILL -- TOUCH ON YESTERDAY'S ASPECTS WITH AN EYE TOWARDS PANCREATIC EYELET REPAIR AND RESTORATION OF HEARING. STAR-- AS FAR AS I CAN TELL I DO NOT SEE ONE OF OUR SPEAKERS. SO A HEAD'S UP DR. JASON SPENT FROM THE UNIVERSITY OF MICHIGAN CAN'T JOIN US BECAUSE OF A LAST MINUTE FAMILY EMERGENCY AND I APOLOGIZE ON THE BEHALF OF THE COMMITTEE AND ANY ORGANIZERS FOR ANY INCONVENIENCE. AS A FRIENDLY REMINDER IT'S TO ENGAGED IN THE NICHD'S IDEA SCALE CAMPAIGN. WHICH WILL BE OPEN THROUGH DECEMBER 16. AND I THINK WE NEED TO ALL IDENTIFY WHAT ARE THE MOST SIGNIFICANT ADVANCES HERE IN THE IDEA SCALE. OUR FIRST SPEAKER IS DR.DOUG MELTON FROM HARVARD UNIVERSITY. DR. MELTON IS THE UNIVERSITY PROFESSOR AT HARVARD MEDICAL SCHOOL AND THE FACULTY OF ARTS AND SCIENCES. HIS RESEARCH FOCUS ON USING CELL AND -- TO MAKE FUNCTIONAL PANCREATIC EYELETS FOR THE STUDY OF DIABETES. HE SO WE WELCOME DR. MELTON. THANK YOU. >> THANK YOU, MARTHA. I WILL NOW TRY TO SHARE MY SCREEN. WITH YOUR PERMISSION. AND ASK YOU TO LET ME KNOW IF THAT IS WORKING. >> CAN YOU NOW SEE -- HOW MANY SLIDES. DO YOU SEE ONE OR SEVERAL? >> WE SEE ONE OF 17 AND YOU'RE IN NOT PRESENTATION MODE YET. >> HOW ABOUT THAT? >> YES. THANKS, DOUG. >> AND THANK YOU FOR INVITING ME TO SPEAK TODAY. I PLAN TO SPEAK ABOUT 15 MINUTES AND IT COMPLIMENTS WHAT I JUST HEARD ON THE BIOPRINTING SECTION IN THAT THE ORGAN OR THE DIABETES CHALLENGE THAT I'M GOING TO SPEAK ABOUT IS SIMPLER IN MANY WAYS BECAUSE IT DOES NOT REQUIRE AS FAR AS I KNOW ANY SPECIAL 3-D STRUCTURE IN ORDER TO PROVIDE FUNCTION. SO AS IT SAYS HERE ON THE SLIDE I'M GOING TO TALK ABOUT USING HUMAN STEM CELLS TO MAKE PANCREATIC EYELETS FOR DIABETES DIABETESES. WHAT I WOULD LIKE TO DO NOW IS BRIEFL-- EXTEND THE DISCOVERY ASPECTS OF LOOKING AT HUMAN STEM CELLS AND FOLLOWING THE CHALLENGE I HAVE A LAST SLIDE WITH FOUR TOPICS FOR LET'S SAY DISCUSSION OR IMPROVEMENT BY THE FIELD WHICH IS JUST MY OWN PERSONAL OPINION. SO FOR THOSE OF YOU WHO DON'T WORK ON DIABETES IT'S A RELATIVELY SIMPLE PROBLEM. THERE ARE ABOUT 100,000 TO 500,000 ORGANOIDS OR ISLETS SHOWN HERE ON THE LEFT. THERE ARE ABOUT 10,000 CELLS AND THE BLUE CELLS ARE THE ONES THAT MAKE THE FAMOUS HORMONE INSULIN. WHAT YOU CAN SEE HERE -- IN TYPE ONE DIABETES THE BETA CELLS ARE DESTROYED BY AN AUTOIMMUNE ATTACK AND TYPE II DYSFUNCTION BY STRESS. WHAT WE SET OUT TO DO IS OBVIOUS. GIVEN THE POTENTIAL OF STEM CELLS. WE SAID INSTEAD OF HAVING PATIENTS MONITOR THEIR BLOOD SUGARS AND INJECT INSULIN AND BE CONCERNED ABOUT THEIR DIET AND EXERCISE EVERY FEW HOURS A DAY. WHY NOT TRY TO DO A NATURAL SOLUTION WHICH IS TO REPLACE THE DYSFUNCTIONAL ISLETS IN TYPE ONE DIABETICS. MANY OF YOU KNOW THIS IS A NONTRIVIAL PROBLEM. WE WERE NO. THE ONLY ONES TO APPROACH THIS AND TO BE HONEST I THOUGHT IT WOULD BE MUCH EASIER. THIS SLIDE HERE SHOWS THE STAGE DIFFERENTIATION PROTOCOL WHICH WE PUBLISHED ABOUT FIVE YEARS AGO. IT'S A SIX STEP PROCEDURE THAT TAKES ABOUT 40 DAYS USING 15 DIFFERENT FACTORS AND I WOULD LIKE TO MAKE A FEW POINTS HERE. ONE, IT WAS MORE DIFFICULT THAN WE THOUGHT. AND IN FACT IN GENERAL MAKING -- ARE MORE DIFFICULT THAN NEURAL DERIVATIVES. TWO, IT WAS VERY CHALLENGING TO GET THROUGH NIH GRANTS EACH OF THESE STEPS ADVANCED BECAUSE THEY DON'T ALL LEAD TO PUBLICATIONS. IMPROVING ONE STEP IS HARDLY AN INTERESTING PUBLICATION. AND IT'S EVEN QUESTIONABLE IF THAT IS A USEFUL FORM OF TRAINING MECHANISMS FOR GRADUATE STUDENTS AND POSTDOCS. AFTER 15 YEARS OF WORK AND I WOULD ESTIMATE ABOUT 50 PEOPLE WE WERE ABLE TO REPLICATE THE NORMAL DEVELOPMENT OF PANCREATIC EYELET CELLS AS SHOWN HERE. THEY ARE STAINED. AND ON THE RIGHT YOU SEE THE STEM-CELL DERIVED STEM CELLS OR ISLETS -- THEY LOOK SIMILAR IF NOT IDENTICAL. I'M GOING TO SHOW YOU TWO SLIDES. ONE OF THEM IS HERE. WHICH IS I COULD SHOW YOU LOTS OF MOLECULAR INFORMATION -. THIS IS THE CHEMISTRY. BUT INSTEAD OF SHOWING THIS ONE BIT OF EVIDENCE -- A HUMAN BETA CELL WITH THE WELL-KNOWN CRYSTALLIZED INSULIN AND ON THE BOTTOM IS THE STEM-CELL DERIVED BETA CELL WHICH HAS THE SAME. AS YOU WILL APPRECIATE IT'S NOT REALLY POSSIBLE TO MAKE THAT KIND OF STRUCTURE UNLESS THESE CELLS HAVE TURNED ON AND OFF ALL THE RIGHT GENES THAT ARE IN THE DEVELOPMENT. NEXT POINT WHICH IS IMPORTANT -- IS THAT THESE CELLS FUNCTION INVITRO AND I'M GOING TO COME BACK TO THAT A FEW TIMES TODAY. THEY DEMONSTRATED PHYSIOLOGICAL FUNCTION FOR THE FIRST TIME. MANY GROUPS HAD SHOWN THAT IT WAS POSSIBLE TO MAKE CELLS THAT FOR EXAMPLE TURN ON THE INSULIN GENE BUT THE CHALLENGE WAS TO MAKE FUNCTIONAL CELLS AND ON THE LEFT PANEL FOLLOWING THREE DIFFERENT GLUCOSE CHALLENGES AT LOW AND THEN LOW AND HIGH AND LOW AND HIGH OR DEPOLARIZATION THE BETA CELLS WE MADE INVITRO MIMIC THOSE THAT WE ISOLATED. THAT I SAID WAS REPORTED IN 2014. THAT WORK WAS ALL DONE IN AN ACADEMIC LAB. AT HARVARD COLLEGE DONE WITH A NONCLINICALLY COMPLIANT CELL LINE SO THE WORK AND THE PROTOCOL WAS TRANSFERRED TO A BIOTECH COMPANY WHICH ADVANCED THE WORK BY IMPROVING THE PROTOCOL BY USING HUMAN EMBRYONIC STEM-CELL LINES. IT HAS REPORTED THE FIRST CLINICAL RESULTS JUST A FEW WEEKS AGO. I'LL SUMMARIZE THOSE AND POINT OUT THAT THEY ARE NOT MY RESULTS BUT IN A SENSE THEY HAVE EFFECTIVELY CURED THE FIRST PATIENT WITH HALF THE CALCULATED DOSE. SO THE PATIENT CLAIMED HE NO LONGER REQUIRES ANY INSULIN. FRANKLY I WAS HOPING THAT THERE WOULD BE SOME DEMONSTRATION OF INSULIN PRODUCTION AND I WAS JUST STUNNED BY THE FACT THAT THIS HALF DOSE OF THE STEM-CELL DERIVED ISLETS PROVIDED TO HIM IN HIS LIVER HAS EFFECTIVELY TAKEN HIM OFF INSULIN AFTER HE HAD BEEN REQUIRING INSULIN FOR 40 YEARS OF HIS LIFE. NOW LET'S REMEMBER THIS IS ONLY THE FIRST HUNDRED DAYS OF THAT PATIENT AND IT'S ONLY ONE PATIENT. BUT I WOULD LIKE TO THINK THAT 100 YEARS AGO WHEN -- THEY INJECTED INSULIN INTO THE FIRST PATIENT AND SAW A RESULT WOULD BE LUCKY ENOUGH TO HAVE A FIRST KIND OF SUCCESS THAT WOULD MIMIC THAT. I'M GOING TO NOW QUICKLY MOVE TO THE NEXT GENERATION NOT DISCOUNTING WHAT HAS BEEN ACHIEVED SO FAR BUT TO FOCUS IN ON HOW THIS PROCESS COULD BE IMPROVED AND THEN POINT TO AREAS THAT I THINK THAT REQUIRE OR DESERVE FURTHER ATTENTION. THE STEM-CELL DERIVED ISLETS ARE HOPE HERE. THEY ARE UNIFORM IN SIZE. THEY CONTAIN ABOUT 10,000 CELLS AS DO HUMAN ISLETS BUT WHEN OUR LAB MAKES THESE CELLS IT'S A RELATIVELY SMALL SET OF CELLS ARE JUST BETA CELLS. HERE IT'S SHOWING 16.5%. TYPICALLY WE MIGHT GET 20% TO 25% BATE AFTER CELLS. I WANT TO TALK ABOUT THE CHALLENGE OF GETTING COMPLETE CONTROL. COMPLETE MASTERY OVER THIS INVITRO DIFFERENTIATION AND THIS IS A CHALLENGE FOR ANY FIELD. I WAS INTERESTED TO LISTEN TO JENNIFER AND OTHERS TALK EARLIER ABOUT TRYING TO MAKE FUNCTIONAL LIVER OR KIDNEY CELLS WHERE YOU HAVE TO MAKE MULTIPLE CELL TYPES AND THEY TO BE IN A CORRECT GEOMETRIC ARRANGEMENT. SO I THOUGHT I WOULD SHARE WHAT WE'VE TRIED AND WHAT HAS NOT WORKED AND THEN MOVE ON TO NEW EXPERIMENTS. ONE OF THE THINGS THAT ONE CAN DO BECAUSE THIS IS ALL DONE INVITRO WITH HUMAN CELLS IS WE CAN ANALYZE EVERY GENE AND EVERY CHANGE THAT OCCURS DURING THE DEVELOPMENT FROM A STEM-CELL ON THE LEFT IN RED TO A FULLY DIFFERENTIATED CELL THE PURPLE ON THE RIGHT OVER HERE. SOMETHING THAT YOU COULD NEVER OBTAIN FROM HUMAN BIOPSIES. WE'VE ANALYZED ALL MODIFICATIONS -- WE HAVE ATTACKED -- TO LOOK AT OPEN CHROMATIN -- AND SIMILARLY USING SINGLE CELL SEQUENCING. WE HAVE ANALYZED ALL OF THE GENES THAT COME ON AND GO OFF. I HAVE TO SAY IN SOME WAYS THIS EXPENSIVE AND TEDIOUS THING IS PROVIDED AN AMAZINGLY DETAILED DESCRIPTION OF HOW A STEM-CELL GIVES RISE TO A BETA CELL AND AN ALPHA CELL. THAT DESCRIPTION MAY BE -- MORE THOROUGH. BUT IT DID NOT GIVE US WHAT I WOULD HOPED. I THOUGHT WE WOULD LEARN ABOUT RECEPTORS THAT APPEAR OR DISAPPEAR AT DIFFERENT STAGES OF THE PROCESSOR SOME CHROMATIN CHANGES WE COULD MANIPULATE BUT WE DIDN'T LEARN ANY OF THAT. WE HAVE A FOUNDATION OF ALL OF THE GENES THAT COME ON AND GO OFF AND ALL OF THE CHROMATIN CHANGES. WHAT I WANT TO FINISH WITH TODAY IS COULD ONE USE GENETIC EDITING TO CONTROL THE COMPOSITION AND PERHAPS MAKE SUPER EYE WILL ITS AND I'LL EXPLAIN WHAT I MEAN BY THAT. * TO BEGIN WITH EMBRYONIC STEM CELLS AND THEN ASK CAN WE RESTRICT DIFFERENTIATION POTENTIAL BY GENETIC EDITING. LIKE CLOSING THE DOOR ON CELL FATE DECISIONS AS THESE CELLS MOVE THROUGH THEIR DIFFERENTIATION. AND COULD WE FIND A WAY TO ADD OR REMOVE GENES THAT WOULD MAKE THESE CELLS MORE IMMUNE COMPATIBLE. SOMETHING MY COLLEAGUE LIKES TO CALL MUNE INERTIA. A WAY TO APPROACH IT. WE GROW OUR CELLS AT ABOUT 100 TO 300 MILLION CELLS IN A STARTING FLASK OF A LITER SHOWN ON THE LEFT. WE INFECT THEM WITH A CRISPR LIBRARY THAT I'LL SHOW YOU IN A MINUTE AND THIS IS INTENDED TO REPRESENT THE SINGLE VIRUS THAT GOT INTO THE CELL. WE DO IT AT A LOW MULTIPLICITY OF INFECTION. NO MORE THAN ONE VIRUS. WE SELECT FOR VIRUSES AND RECOVER THE CELLS AND SEED THEM AND GO THROUGH THE 40 DAY DIFFERENTIATION PROCEDURE. FIX THE CELLS AND STAIN FOR MARKERS FOR ALPHA, BETA AND OTHER UNWANTED CELL TYPES UNDER THE FIXATION AND SEQUENCE THE GUIDE DNAs. LIBRARY WE USED WE OBTAINED FROM THE BRODE INSTITUTE. WE'VE DONE THIS IN A THOROUGH WAY. WE SCREENED THE ENTIRE HUMAN GENOME. AND THEN WE SELECTED ABOUT 800 CANDIDATES. WE DID 9,000 TOTAL -- NONTARGETTING. AND THIS TOOK US ABOUT TWO YEARS TO WORK OUT HOW TO DO THIS BUT I THINK THIS APPROACH IS NOW APPLICABLE TO A STUDY OF ANY DIFFERENTIAL CELL TYPE AND ASK WHICH GENES YOU MIGHT WANT TO TURN ON AND OFF TO CHANGE THE RESULTS. WITH THIS NEXT SLIDE HOW THE DATA HAS COME OUT AND WHAT WE'RE DOING WITH IT. I'M GOING TO GIVE YOU TWO EXAMPLES AND I'M SHOWING YOU THE SAME DATA IN TWO FORMATS. THE FIRST FORMAT IS TO LOOK ON THE LEFT AT THIS BAR DIAGRAM WHERE EACH DOT REPRESENTS A KNOCKOUT. THIS HAS TO DO WITH THE STATISTICAL SIGNIFICANCE IN THE POPULATION AND THE POINT WOULD BE IF WE KNOCKOUT THE TRANSCRIPTION FACT KNOWN TO BE REQUIRED FOR BETA CELL DEVELOPMENT YOU SEE KNOCKING THAT CHAIN OUT MAKES IT MUCH LESS LIKELY TO BECOME A BETA CELL. HERE IT IS SHOWN IN A BUY DIAGRAM WHERE THE X IS OPPOSITE SC BETA. IF I CAN DRAW YOUR ATTENTION TO THE LOWER RIGHT -- OTHER LABS HAVE SHOWN THAT -- ARE REQUIRED FOR ALPHA CELLS AND WHEN IT'S KNOCKED OUT YOU'RE LESS LIKELY TO BECOME AN ALPHA CELL. WE DON'T HAVE MONOCLONAL CELLS. TO KNOCKOUT THESE GENES HERE WHICH WILL MAKE YOU LESS LIKELY TO BECOME THE CELL TYPE WE DON'T WANT. AND TO KNOCKOUT THESE GENES HERE WHICH MAKE IT MORE LIKELY THAT YOU WILL BE BETA ORAL FROM CELLS. I HOPE THAT IS COMING IN THE NEXT YEAR. I'M GOING TO FINISH NOW -- BY SAYING SOMETHING -- MAYBE A LITTLE BIT MORE DREAM LIKE WHICH IS UP TO THE LAST FEW YEARS MY LAB FOCUSED ON TRYING TO MAKE NORMAL OR NATURAL ISLETS AND I'VE DECIDED TO MAKE SOMETHING THAT IS LIKE A SUPER EYELET. THAT MIGHT NOT BE BOTHERED OR MIGHTY SATISFIED THE IMMUNE SYSTEM. * SO MYAMAR VERY WELL TRICK IS TO USE A MARVEL SHIELD TO SAY WITH DEGENETICALLY MODIFY THE CELL TO THEY ARE NOT EASILY IDENTIFIED AND REJECTED WHEN TRANSPLANT INTO PATIENTS. THESE ARE THE GENES WE'VE KNOCKED OUT. MANY ARE OBVIOUS. FOR EXAMPLE IF YOU LOOK AT IMMUNE MODULATION IT'S OBVIOUS NOW WITH WORK IN CANCER I MOON OHIO THERAPY. AND THE CRISPR SCREENING THAT I'VE DESCRIBED. I WANT TO FINISH THEN WITH JUST MY OWN PERSONAL VIEW * ON WHERE THE FIELD OF TREATING BIBB WITH CELL TRANSPLANTATION WILL GO AND THEN TAKE UP THE CHALLENGE ON WHAT I THINK IS MISSING. SO WHAT IS ALREADY HAPPENED NOW IS TRANSPLANTATION INTO PATIENTS -- THESE ARE STEM-CELL DERIVED ISLETS USING HUMAN EMBREEIANIC STEM CELLS. WE'LL SEE THOSE CELLS PUT IN -- TRANSPLANTATION AND I WOULD LIKE TO DREAM ABOUT A TIME WHEN AN ENDO VERY NOT JUST HAVE IN HER FREEZER NAKED GENETICALLY MODIFIED CELLS. THEY ARE TOLD THEIR CHILD HAS TYPE ONE DIABETES. SHE SAID THAT IS NOT A BIG PROBLEM. WE HAVE THESE CELLS THAT WILL NOT BE REJECTED AND SQUIRT OUT THE RIGHT AMOUNT OF INSULIN AND THAT WOULD BE RIGHTLY CALLED A CURE FOR THE DISEASE. I WILL FINISH BECAUSE I'M TRYING TO STAY ON TIME. FOUR TOPICS THAT I THOUGHT WE MIGHT WANT TO DISCUSS. EMPHASIZE THESE ARE MY OWN OPINIONS. THE FIRST CHALLENGE IN THE FIELD I BELIEVE IS THAT THERE ARE ONLY A HANDFUL OF FUNCTIONAL CELLS THAT HAVE BEEN MADE FROM HUMAN STEM CELLS. NEURONS, -- MAYBE SOME LIVER CELLS. BUT MAKING THE KIDNEY, MAKING OTHER KINDS OF NEURONS IS PROVEN TO BE VERY DIFFICULT AND THE LITERATURE IS FULL OF CLAIMS WHICH ARE UNSUBSTANTIATED. BECAUSE A CELL MAKES INSULIN THAT DOESN'T MAKE IT A BETA CELL. BECAUSE A CELL MAKES -- THAT DOESN'T MAKE IT A LIVER CELL. SO IT'S A CHALLENGE TO BOTH GET FUNCTIONAL CELLS AND TO GET THEM FROM CLINICALLY COMPLIANT HUMAN STEM CELLS. THE SECOND POINT AND THIS HAS BEEN DISCUSSED IS THAT SCALING UP AND MANUFACTURING PRESENTS ENORMOUS CHALLENGES AND I WOULD SAY OPPORTUNITY FOR INNOVATIVE. THERE ARE HARDLY ANY ACADEMIC LABS THAT ARE PREPARED TO TAKE ON THE SCALE UP PROCESS AND FROM WHAT I KNOW IN THE INDUSTRY WHICH DOES THIS STUFF THEY ARE USING AGE OLD METHODS THAT ARE RIPE FOR INNOVATIVE. SO IF I WERE IN CHARGE I WOULD SAY LET'S TRY TO ENGAGE MORE BIOENGINEERS AND CHEMICAL ENGINEERS IN HOW TO DEAL WITH SCALE UP AND MANUFACTURING AND THEN AS WE HEARD IN THE LAST SESSION EVENTUALLY ORGANS. THIRD POINT I WOULD MAKE IS THAT I PERMANENTLY BELIEVE NOW THE BIGGEST CHALLENGE IS HOW TO INDUCE TOLERANCE AND PROTECTION. FOR STEM-CELL DERIVED TISSUES. I DO NOT BELIEVE THERE WILL BE A TIME AT LEAST IN MY LIFETIME WHEN PEOPLE WILL BE MAKING PERSONALIZED IPS CELLS FOR INJECTION. IT'S TOO EXPENSIVE. EACH ONE WOULD REQUIRE ENORMOUS COST TO MAKE THE LINE AND FIGURE OUT HOW TO DIFFERENTIATE IT. I DON'T THINK THAT IS COMMERCIALLY FEASIBLE. THEREFORE I BELIEVE ONE SHOULD FOCUS IN ON REJECTION TO TRY TO FIND WAYS OF IMMUNE PROTECTION AND TOLERANCE. THIS OBVIOUSLY INVOLVES SOMETHING FAR OUT OF MY AREA OF EXPERTISE. WE HAVE AN ACTIVE IMMUNOLOGY COMMUNITY WORKING ON ALL ASPECTS OF THIS. AND I THINK THAT INFORMATION WILL BE GREATLY INFORMATIVE FOR STEM-CELL DERIVED CELL TRANSPLANTS. AND LASTLY A PITCH FOR WHAT I THINK IS ALWAYS TRUE. WE DON'T HAVE GOOD MODELS. I DON'T MEAN TO SAY THAT THE HUMANIZED MICE ARE NOT INFORMATIVE BUT THEY ARE NOT VERY GOOD MODELS FOR THE FIELD THAT I WORK IN WHICH IS HOW WOULD YOU TRANSPLANT -- DERIVED STEM-CELL DERIVED ISLETS INTO A PATIENT. YOU CAN DEMONSTRATE REJECTION BUT THEY ARE NOT SUFFICIENTLY INDICATIVE OF WHAT HAPPENS IN HUMANS. SO I WOULD LIKE TO MAKE A CALL FOR NONHUMAN PRIMATES AS THE TISSUES COME TO THE FLOOR. I'LL FINISH BY POINTING OUT THAT PEOPLE THAT HAVE DONE THIS WORK IN MY LAB -- ON THE LEFT -- ON THE BOTTOM OF THE FOUNDATION THAT'S HAVE PROVIDED FUNDING IN GREEN IS OUR STEM-CELL FOUNDRY. AND IN BLUE ARE THE COLLABORATORS. I'M SORRY I MAY HAVE GONE OVER A LITTLE BIT BUT I'M GOING TO SEND THIS SHOW NOW AND SEE IF I CAN STOP SHARING. YES. STOP SHARING SO MARTHA SORRY FOREGOING OVER >> NO PROBLEM. IT IS GREAT TO HAVE YOU ON HERE. VERY EXCITING WORK. I HAVE A COUPLE OF QUESTIONS FOR YOU AFTER OUR NEXT SPEAKER. 7 SO WE'LL HOLD -- HAVING QUESTIONS UNTIL THE END. DR. JASON SPENCE FROM THE UNIVERSITY OF MICHIGAN WILL NOT BE ABLE TO JOIN US TODAY BECAUSE OF A LOCAL FAMILY EMERGENCY BUT I DO WANT TO INTRODUCE OUR ESTEEMED NEXT SPEAKER. DR. HERTZANO WHO'S A PROFESSOR IN -- HEAD AND NECK SURGERY. SHE GOT HER TRAINING IN TEL AVIV AND CURRENTLY AT THE UNIVERSITY OF MARYLAND IN THE SCHOOL OF MEDICINE IN BALTIMORE. HER CLINICAL PRACTICE FOCUSES ON THE DIAGNOSIS AND TREATMENT OF DISEASES OF THE EAR WITH AN EMPHASIS ON HEARING RESTORATION. AND TOWARDS HEARING RESTORATION SHE HAS LED A COLLABORATIVE TEAM THAT DEVELOPS AND APPLIES A VARIETY OF APPROACHES FOR CELL TYPE SPECIFIC MULTI-OPENEDIC ANALYSIS OF THE EAR. THEY COUPLE SOME OF THE RESULTS WITH STATE-OF-THE-ART INFORMATICS AND ANALYSIS TO SPECIFIC SIGNALING CASCADES THAT AN A INQUIRED HEARING LOSS. HER WORK HAS LED TO THE INCEPTION AND DEVELOPMENT OF G EAR A PORTAL FOR GENE EXPRESSION FOR ANALYSIS RESOURCE SHARING AND IT'S A PORTAL FOR INTUITIVE VISUALIZATION AND ANALYSIS AND SHARING OF GENE EXPRESSION DATA AND WE WELCOME DR. HERT WERE, ANO TODAY. THANK YOU. 7 * THANK YOU, MARTHA FOR THE VERY KIND INTRODUCTION. I WILL SHARE MY SCREEN. I HOPE YOU CAN SEE MY SCREEN. >> YES. LOOKS GREAT. >> I'M A SURGEON SCIENTISTS. AND MY PRACTICE FOCUSES ON HEARING RESTORATION. AND OUR WORK WITH REGARDS TO REGENERATION REALLY FOCUSES ON FINDING WHAT ARE THE FACTORS THAT ARE NEEDED TO ADD TO THE MIX. SO I'M ENVIOUS WHEN I HEAR THE PREVIOUS SPEAKERS AND THE FACTORS AND -- THAT ARE ALREADY THERE FOR THE MOST PART AND IN THE EAR FIELD WE'RE CERTAINLY NOT THERE. HAVE ONE DISCLOSURE. AND FOR THOSE OF YOU WHO ARE NOT FAMILIAR WITH THE FIELD AS A REMINDER -- OUR SENSE OF HEARING IS AS TRULY REMARKABLE. SOUNDS TRAVELS BY AIR. THE ENERGY IS FOCUSED FROM THE RELATIVELY VERY LARGE -- MEMBRANE ON TO THE SMALL PLATE RESULTING IN A FLOYD WAVE THAT TRAVELS THROUGHOUT THE ORGAN OF THE HEARING WHICH IS CALLED THE COCHLEA AND RESULTS IN VIBRATION OF THE SENSORY CELLS THAT ARE THROUGHOUT THE LENGTH OF THE COCHLEAR DUCT. THEY ARE CALLED HAIR CELLS BECAUSE OF THE ASK TIN -- -- THESE ARE CALLED -- AT THE TIPS HAVE A MECHANICAL CHANNEL. THE MECHANICAL ELECTRICAL TRANSITION CHANNEL. THEN THERE IS AN ACTION POTENTIAL AND THIS MECHANICAL FORCE OF SOUND IS TURNED INTO AN ELECTRICAL IMPULSE TRAVELS THROUGH THE NERVES TO THE BRAIN. THE SENSORY CELLS ARE ORGANIZED IN FOUR ROWS. A ROW OF INNER HAIR CELLS AND THREE ROWS OF OUTER HAIR CELLS AND THIS IS A GRAPH FROM A MOUSE HERE AND WHEN WE'RE BORN IF WE DON'T HAVE ANY GENETIC DEFECTS WE HAVE A FULL COMPLIMENT OF THESE THAT RUN THROUGHOUT THE LENGTH OF THE COCHLEA AND THE HEARING THRESHOLDS OF PEOPLE AGE 10-21 FAR EXCEED WHAT WE NEED FOR SPEECH AND SPEECH IS THE MOST IMPORTANT THING. COMMUNICATION IS CERTAINLY THE MOST IMPORTANT THING THAT WE DO WITH OUR SENSE OF HEARING. 7 SO YOU CAN SEE THAT EARLY ON WE CAN HARRIESLY FROM 125 Hz TO ALMOST 20,000 Hz. BUT WITH NORMAL HEALTHY AGING EVEN BY AGE 60 WE LOSE MANY OF THESE FREQUENCIES AND THE THRESHOLD TO HEAR MANY OF THESE FREQUENCIES INCREASES DRAMATICALLY LARGELY BECAUSE OF A LOSS OF THE SENSORY CELLS -- AND THEY DO NOT REGENERATE. THIS IS SO COMMON THAT IF YOU GOOGLE HOW OLD ARE MY EARS YOU WILL FIND YOUR AGE. THIS IS ALSO SO COMMON THAT ACCORDING TO MARCH 2021WHO STATEMENT THERE ARE 430 MILLION PEOPLE WORLDWIDE WITH DISABLING HEARING LOSS AND THE NUMBER IS GOING TO INCREASE A LOT OF WHICH IS SELF-INFLICTED FROM NOISE EXPOSURE. SO GENERALLY SPEAKING HOW DO WE TREAT HEARING LOSS? SADLY NOT BASED ON THE CAUSE OF THE HEARING. WE BASICALLY TREAT HEARING LOSS BASED ON THE DEGREE OF THE HEARING LOSS. LITTLE HEARING LOSS IS TREATED WITH HEARING AIDS AND MORE HEARING LOSS IS TREATED WITH COCHLEAR IMPLANTS AND IN OUR TEAM WE TRY TO FIGURE OUT AND IDENTIFY FACTORS FOR HAIR CELL REGENERATION. AND ALSO WE WORK SEPARATELY AND A DIFFERENT PORTFOLIO I'M NOT GOING TO TALK ABOUT TODAY BUT IDENTIFYING COMPOUNDS FOR OTO PROTECTION. IT'S EASIER TO PROTECT SOMETHING THAN TO REBUILD IT ONCE IT IS LOST. SO TOWARDS -- REGENERATION WE WERE ASKING OURSELVES WHAT ARE THE TRANSCRIPTION FACTORS THAT ARE NECESSARY TO FORM A HAIR CELL. AND BACK DURING MY GRADUATE DEGREE IDENTIFIED THAT THE TRANSCRIPTION FACTOR -- TURNS ON GF I1 AND -- -- THEY ARE REALLY CONSIDERED AS THE -- FACTORS OF HAIR CELLS. AND THE PROBLEM IS WHEN YOU TAKE ALL THREE OF THEM AND PUT THEM IN A STEM-CELL YOU GET SOMETHING THAT LOOKS LIKE THIS AND IT'S A FAR CRY FROM A REAL MATURE HAIR CELL SO WE WERE ASKING OURSELVES HOW CAN WE GO AND LOOK AT DIFFERENT REGULATORS. HOW CAN WE IDENTIFY MORE REGULATORS. IT WAS IDENTIFIED BY DOING A MICRO RAY AND IDENTIFYING GENES THAT ARE MISS REGULATED BUT THAT IS NOT ENOUGH. 8 SO A LOT OF MY WORK IS DONE WITH A MATHEMATICIAN. WE'VE BEEN WORKING TOGETHER SINCE WE WERE GRADUATE STUDENTS AND THE IDEA THAT WE HAD WAS IF WE TAKE DIFFERENT CELLS AND THE SAME CELL AND DIFFERENT TIME POINTS -- AND WE LOOK AT THEIR GENE EXPRESSION WE FIND THAT MOST OF THE GENES ARE COMMON BUT THEY ARE UNIQUE GENES. FOR CELL TYPES YOU CAN IMAGINE THAT A HAIR CELL HAS UNIQUE GENES. THEY HAVE DIFFERENT PROGRAMS THAT THEY NEED TO ACTIVATE. AND IF WE LOOK AT THE REGULATORY REGIONS OF THE GENES THAT ARE UNIQUELY EXPRESSED AT THE STATE OR CELL TYPE WE WOULD EXPECT TO FIND THAT THE CELL TYPE OR STATE SPECIFIC GENES WILL HAVE SOME ENRICHMENTS FOR SHORT SEQUENCES THAT WOULD END UP BEING THE DOCKING STATIONS OR THE RECOGNITION SITES FOR TRANSCRIPTION FACTORS. SO THAT WAS A NICE IDEA. HOWEVER THE INNER EAR IS VERY COMPLEX. THIS IS A CROSS SECTION. HERE ARE THE NEURONS. YOU CAN SEE THE CROSS SECTION THERE ARE ONLY FOUR HAIR CELLS AND TENS OF THOUSANDS OF CELLS. SO JUST EXTRACTING RNA FROM THE COCHLEA IS NOT GOING TO CUT IT. AND INITIALLY THAT WAS BEFORE SINGLE CELL -- WERE AVAILABLE. WHAT WE DID WAS WE FIGURED THAT SOME OF THE CELL SURFACE MARKERS USED BY IMMUNOLOGIST MAY ALSO BE EXPRESSED IN THE EAR. WE FOUND THAT EACH ONE OF THEM HAS A DIFFERENT PATTERN OF EXPRESSION. 7 SO THIS ALLOWED US TO PUT TOGETHER A PROTOCOL THAT WAS ESSENTIALLY A PUZZLE THAT ALLOWED US TO BREAK DOWN THE EAR -- AND YOU CAN SEE WHEN WE USED THESE ANTIBODIES AT STANDARD CONCENTRATIONS WE GET A NICE SEPARATION TO POPULATIONS WHICH ALLOWED US TO EXTRACT RNA AND ASK THE QUESTION. ARE THERE GENES THAT ARE UNIQUELY EXPRESSED IN THESE DIFFERENT CELL TYPES OF THE YEAR AND OUR FIRST TRY WAS LOOKING AT -- ESPECIALLY THINKIAL -- * AND YOU CAN SEE THERE ARE LARGE GROUPS OF GENES THAT WERE UNIQUELY EXPRESSED. AND THEN ASK IS THERE A SHORT SEQUENCE THAT IS OVER REPRESENTED IN THOSE CELL TYPES SPECIFIC GENES. AND INDEED WE FOUND A SHORT SEQUENCE THAT WAS OVER REPRESENTED WITH A HIGH P VALUE. THIS ENDED UP BEING A PERFECT MATCH TO THE BINDING SITE OF THE TRANSCRIPTION FACTORS WHICH IS A KNOWN REGULATOR OF -- FATE AND OTHER SYSTEMS AND WE HAD A COLLEAGUE THAT HAD A MOUSE WITH A MUTATION AND THE INNER EAR WAS HIGHLY DEFORMED. SO IT WAS ESPECIALLY THINKIAL FATE. * THAT ALLOWED US TO MOVE ON TO HAIR CELLS AND THE QUESTION WAS AT THE NEWBORN STATE CAN WE USE THE SAME APPROACH TO IDENTIFY A TERMINAL REGULATOR DIFFERENTIATION. YOU GET IMMATURE HAIR CELLS SO WE COMBINED THE PROTOCOL WITH A MOUSE THAT EXPRESSES A GREEN FLUORESCENT PROTEIN. THAT ALLOWED US TO ISOLATE VERY PURE POPULATIONS. AND DO AN RNA SEEK. AND HERE AGAIN IN THE -- HAIR CELL SPECIFIC GENES WE IDENTIFIED AN ENRICHMENT WITH A VERY GOOD 3, VALUE OF A BINDING SITE OF TRANSCRIPTION FACTOR. AND THIS WAS DIFFERENT THAN THE TRANSCRIPTION FACTOR THAT WAS COMMON -- A VERY DIFFERENT SIGNATURE AND HOWEVER THERE WERE 8 TRANSCRIPTION FACTORS THAT MATCHED BECAUSE -- THERE WERE 8 TRANSCRIPTION FACTORS SO WE NEEDED TO FIND A WAY TO CONVINCE OURSELVES. WE REPEATED THESE ANALYSIS -- WE WERE QUITE ASTONISHED TO FIND THE VERY SAME SIGNATURE. AND THIS SUGGESTED THAT THE RFX TRANSCRIPTION FACTORS ARE EVOLUTIONARY CONSERVED REGULATORS OF CELL FATE IN HAIR CELTS AND WE DECIDED TO MAKE THE INVESTMENT AND MAKE THE ADDITIONAL KNOCKOUT FOR ALL OF THEM AND TRY TO SEE WHAT HAPPENS WHEN WE DON'T HAVE R EXPECTS TRANSCRIPTION FACTORS. THIS WAS A LONG PROJECT BECAUSE AS YOU SEE IN THESE GRAPHS WHICH ARE HEARING CHARTS LOOKING AT THE SENSITIVITY AT THREE DIFFERENT FREQUENCIES IN BLACK ARE THE -- -- THE MUTANTS HAD THE SAME HEALING THRESHOLDS SO WE STARTED ASSUMING REDONE SEE AND 3 AND 7 HAD THE HIGHEST LEVEL OF EXPRESSION BUT THEY ALSO HAD NORMAL HEARING THRESHOLDS. WITHIN ONE AND THREE WERE FROM THE SAME FAMILY AND WE HAD A SEVERE HEARING LOSS. AND WE TOOK THESE MICE OUT TO THE MICROSCOPE. YOU CAN SEE THE HAIR CELLS AND PARTICULARLY THE OUTER HAIR CELLS ARE MISSING AND THE INNER HAIR CELLS ARE DEGENERATING. ISSUE HERE WAS THAT THE TISSUE WAS FROM 3-MONTH-OLD MICE AND THE ANALYSIS WAS ORIGINALLY ON ONE DAY OLD MICE. NOW LOOKING FOR DIFFERENT FACTORS AND THAT SUGGESTED THAT WE MISSED THE PARTY SO WE TOOK MORE MICE AND STARTED DOING FUNCTIONAL ANALYSIS AND SAW THAT EARLY ON THREE DAYS AFTERBIRTH THE TRANS ACCEPTS CHANNEL IS THERE *. SOME OF THE PROTEINS -- ARE THERE. 7 AND ULTRA STRUCTURALLY UP TO 8 DAYS THINGS LOOK GOOD. IN MICE ON SET OF HEARING IS ONE THE MICE ARE AROUND 13 DAYS. SO WE LOOKED UP TO 12 DAYS AND WE DIDN'T SEE ANY DIFFERENCE. BUT JUST THREE DAYS AFTER THE ONSET OF HEARING YOU CAN SEE ALL OF THE HAIR CELLS ARE LOST. SO THAT LED US TO THE QUESTION ARE THESE FACTORS -- OF SURVIVAL AND MUCH MORE WORK THAT WAS DONE SHOWS THROUGH CONDITIONAL DELETION OF TIME POINTS THESE ARE TERMAL DEPRECIATION FACTORS -- JUST TO SHOW ANOTHER EXAMPLE AND HOW THIS METHOD REALLY WORKS AND OUR NEXT QUESTION WAS THE OUTER HAIR CELLS THEMSELVES BECAUSE SINCE THE 1800s WE KNOW THAT INNER OR OUTER -- WHEN YOU TRY TO TURN THEM INTO AIR CELLS WHAT DON'T GET IS OUTER HAIR CELLS. HOW CAN YOU FIGURE OUT AND GET THIS HIGHLY SPECIALIZED CELL TYPE FOR THE OUTER HAIR CELLS. THEY ARE VERY SENSITIVE AND ASSOCIATING THEM WITH NUMBERS IS DIFFICULT WITHOUT CHANGING -- SO WE USED A MODEL THAT ALLOWED US TO -- -- -- -- AND OUR ANALYSIS POINTED TO FIVE CANDIDATE TRANSCRIPTION FACTORS AND AS OFTEN HAPPENS WE WERE LUCKY. BECAUSE ANOTHER GROUP FROM THE UK THAT WAS INVOLVED IN THE SCREENING FOR AGE RELATED HEARING LOSS IDENTIFIED A MOUSE MODEL THAT HAD THE FULL COMPLIMENT OF HAIR CELLS BUT THE MOUSE WAS DEAF. SO THEY CONTACTED US AND SHOWED US THE STAINING OF THIS TRANSCRIPTION FACTOR -- THAT WAS REALLY UNIQUELY AND BEAUTIFULLY EXPRESSED ONLY IN THE OUTER HAIR CELLS. 8 AND HERE IS THEIR FUNCTIONAL DATA. SO REALLY OUR QUESTION AS REGULATORS OF GENE DIFFERENTIATIONS THIS REALLY WHAT GOVERNS REGULATION OF GENE EXPRESSION BETWEEN INNER HAIR CELLS AND OUTER HAIR CELLS AND CAN THIS BE USED TO DRIVE THE GENERATION OF STEM CELLS. WE TOOK MICE THAT HAVE A RED FLUORESCENT PROTEIN IN ALL OF THE HAIR CELLS. WE PUT THE -- OR FGP GENE -- AND WE INJECTED IT INTO THE INNER EARS OF NEWBORN MICE. WE HAVE WAITED A FEW DAYS AND COLLECTED THE CELLS. AND FROM THE CONTROLS WE WERE ABLE TO DEVELOP A SET OF MARKERS FOR THE INNER AND OUTER HAIR CELLS AND WE COLLECTED CELLS FROM THE TRANSCRIPTION FACTOR. AND OUR EXPECTATION WAS THAT IF THIS IS INDEED A MASTER REGULATOR THEN THE INNER HAIR CELLS MAY TURN OFF SOME OF THE PROGRAMS. THE OUTER HAIR CELLS WE DID NOT KNOW WHAT WOULD HAPPEN. SO LET'S SEE WHAT HAPPENED. THE INNER HAIR CELL -- HAVE INNER HAIR CELL MARKER GENES. AND YOU CAN SEE -- HERE THOSE GENES GO DOWN. SIMILARLY WHEN WE TAKE OUTER HAIR CELL GENES THAT ARE EXPRESSED THESE START TO BE TURNED ON. AND WHEN WE LOOK AT THE PROTEIN LEVEL AND PReS IN TO THE HALLMARK MOLECULE OF OUTER HAIR CELLS NORMALLY EXPRESSED ONLY IN THE THREE ROWS OF THE OUTER HAIR CELLS. THE INNER HAIR CELLS EXPRESS IT TOO. THEY ALSO ACQUIRE -- WHICH IS THE HALLMARK FEATURE OF OUTER HAIR CELLS. THIS PROJECT IS CONTINUED BY CHRISTOPHER IN MY LAB AND I WILL SAY THAT EVEN THOUGH THE RESULTS ARE AMAZING -- IT DOES NOT REGULATE THE -- SURFACE AND WHEN WE WERE ESTIMATING FOUR CRITICAL PROGRAMS FOR OUTER HAIR CELLS REGULATING ONE OF THEM AND WE'RE LOOKING FOR THE OTHER REGULATORS. SO WE APPLIED THIS FOR MANY OTHER PROJECTS AND THIS WORKS. SO CERTAINLY SOMETHING THAT CAN BE TRANSFERRED TO OTHER SYSTEMS. AS WE'RE DOING ALL OF THIS WORK WE'RE GENERATING A LOT OF OMIC DATA AND WE WANTED TO BE ABLE TO SHARE THIS DATA IN A MEANINGFUL WAY WITH OTHER LABS AND WE WANTED TO HAVE ACCESS TO DATA FROM OTHER LABS AND WE DIDN'T KNOW HOW TO DO IT UNTIL WE CAME UP WITH THE IDEA OF PUTTING TO THE SOMETHING THAT IS CALLED THE EAR OR GENE EXPRESSION ANALYSIS RESOURCE. AND IT BASICALLY GIVES ASSETS TO OMIC DATA AT LEVEL AND EASE THAT WE HAVE ACCESS TO READING MANUSCRIPTS IN PubMed. YOU NEED TO TAKE A TYPEWRITER AND WRITE THE MANUSCRIPT AGAIN BUT WHEN WE HAVE OMIC DATA YOU NEED TO DOWNLOAD IT AND WHAT IS DEPOSITED IT IS THE RAW READ WHICH IS USED SEVEN YEARS BEFORE PUBLICATION. SO YOU PUT IN ONE OR MORE GENES AND IT PRESENTS MULTIPLE DATASETS WE HAVE FOR THE EAR FIELD. WE HAVE OVER 150 OF THOSE ORGANIZED IN THE PROFILE. THEY ARE PRESENTED IN MANY DIFFERENT WAYS FROM -- FOR SINGLE CELL TO GRAPHICAL CARTOONS. THAT GENE EXPRESSION IS COLORIZED. BAR GRAPHS AND YOU CAN CURATE AND CHANGE THE PRESENTATION YOURSELF THERE IS INFORMATION ABOUT WHETHER ANY GENE YOU'RE LOOKING AT -- -- -- -- AND YOU CAN CHANGE THE DISPLAY. YOU CAN ANALYZE THE DATA WITHOUT DOWNLOADING IT. JUST PReS AND COMPARE. AND YOU CAN GO INTO THE SINGLE CELL YOU CAN ANALYZE THE NOVA. THERE IS A LOT THERE AND THE NEAT THING IS THAT THIS CAN BE REPURPOSED TO ANY FIELD. FOR DIABETES. FOR EYE. WE HAVE RECENTLY REPURPOSED IT FOR BRAIN INITIATIVE AND IT IS THE OFFICIAL -- ANALYSIS FOR BRAIN INITIATIVE. SO FOR THOSE OF YOU THAT ARE INTERESTED. WE'RE ALWAYS LOOKING FOR CHAMPIONS TO TAKE IT FORWARD FOR OTHER FIELDS. SO WITH THIS I DON'T WANT TO RUN OVER. I'LL JUST THANK MY FUNDING RESOURCES. WE'VE GOT GENEROUS FUNDING FOR THE NIH. AND FOUNDATIONS. 8 AND MY TEAM MEMBERS AND I'M HAPPY TO TAKE QUESTIONS. >> THANK YOU, RONNA. THAT WAS AN EXCITING TALK AND THIS IS WAY OUTSIDE MY FIELD. A CARDIOVASCULAR MEDICINE BASED. I HAVE A REQUEST ABOUT YOUR AUTO SORT TECHNOLOGY. THE SPECIFIC GENES THAT ARE UNIQUE TO THE SENSORY ESPECIALLY THINK YAM. AND YOU MENTIONED THAT TRANSCRIPTION FACTORS -- ARE EVOLUTIONARY -- IN TERMS OF THE RFX GENES AND THAT YOU ACKNOWLEDGE THAT MAYBE YOU MIGHT HAVE MISSED THE PARTY USING THE MOUSE MODEL. I'M ASKING A QUESTION HOW MUCH ENVIRONMENT PLAY A ROLE IN TERMS OF THE DIFFERENTIATION OF OUTER VERSUS INNER HAIR CELLS? -- I COULD TAKE THE INNER EAR AND YOU GROW IT EVEN INVITRO. SO TAKE AN E12 WHICH DOES NOT HAVE HAIR CELLS AT ALL AND YOU JUST GROW IT IN A CULTURE DISH AS ANNEX PLANT. YOU WILL GET FULL DIFFERENTIATION. >> SO I GUESS TO ASK A LITTLE BIT DEEPER. SO ENVIRONMENTAL NOISE. DOES THIS IMPACT HOW THE OUTER VERSUS THE INNER HAIR CELLS DEVELOP AND MAINTAIN THEIR STATUS? >> SO THERE IT DOES NOT EFFECT DEVELOPMENT TO OUR KNOWLEDGE. BUT OUTER HAIR CELLS ARE MUCH MORE VULNERABLE AND THE THE REASON THAT WE LOSE OUR HEARING WITH AGE AND WE ALL DO. TO BE HUMAN IS TO LOSE YOUR HEARING. IS BECAUSE THE HIGHER FREQUENCIES. THE HAIR CELLS ARE THE BASE OF THE COCHLEA WHERE SOUND COMES IN AND EVERY DAY WE LOSE OUTER HAIR CELLS ALL THE TIME. AND UNFORTUNATELY THEY DO NOT REGENERATE. BUT -- THEY -- -- THEY DO REGENERATE SO A BIG PART OF THE HRP PROJECT THAT I'M PART OF IS COMPARING THE RESPONSE TO DAMAGE AS YOU SUGGEST. BETWEEN SPECIES THAT DO AND DO NOT REGENERATE HAIR CELLS AND TRYING TO FIND THE CUES THERE. >> AWESOME. SO IN THE SAME VEIN DOUG I HAVE A QUESTION ABOUT EYELET CELLS WITH REGARDS TO TYPE ONE DIABETES. ISLETS. -- AND I JUST WONDERED IS THERE ANYTHING THAT WE CAN LOOK AT IN TERMS OF HOW MUCH DOES THIS APPLY TO THE PDX IS AND INS1 PROFILES IN TERMS OF EXERCISE AND ALSO IN TERMS OF ALOE VERSUS AUTOIMMUNITY IS THERE ANYTHING THAT WE SHOULD KNOW ABOUT INDIVIDUALS EITHER TYPE ONE OR TYPE II AND WHERE SOME PEOPLE HAVE DIFFERENT EXERCISE REGIMENS VERSUS NOT. >> WELL THANKS, MARTHA AMOUNT A COUPLE OF QUESTIONS. FIRST I'LL POINT OUT THAT FOR TYPE ONE DIABETES YOU HAVE BOTH THE ALOE AND AUTOIMMUNE RESPONSE SO YOU HAVE TO DEAL WITH BOTH OF THEM. FOR TYPE II YOU HAVE ORIGINAL AN ALOE RESPONSE. THAT IS WHY I THINK DOING GENETIC SCREENINGS I'M TRYING TO REDUCE REJECTION GOING FORWARD. ABOUT EXERCISE -- FOR PEOPLE THAT HAVE RECEIVED ISLET TRANSPLANTS ALONG WITH IMMUNO SUPPRESSION HAVE BEEN OFF SINCE LYNN FOR 20 YEARS AND IN MANY CASES AND THEY DON'T HAVE ANY SPECIAL EXERCISE REGIMEN OR DEFICIENCIES OR REQUIREMENTS. THAT IS WHAT I MEANT BY PROVIDING NATURAL BETA CELLS YOU ARE DEALING WITH BOTH CONTROLLING BLOOD SUGARS BY READING BLOOD SUGAR LEVELS EVERY MILLISECOND AS OPPOSE TO FIVE MINUTES AND YOU'RE JUST PUTTING OUT TINY BITS OF INSULIN ALL THE TIME. SO THE BLOOD SUGARS ARE CONTROLLED NORMALLY BY ISLET TRANSPLANTS. >> AWESOME. THAT IS A GREAT ANSWER. I WOULD LIKE TO ALSO GET TO MAYBE THE AUDIENCE HAS SOME SPECIFIC QUESTIONS. BUT I HAVE OTHER GENERAL QUESTIONS JUST TO ASK YOU BOTH. IF YOU DON'T MIND. >> ONE THING FORE THE FIELD FOR BOTH HEARING AND DIABETES MANAGEMENT WHAT ARE THE SPECIFIC IMPEDIMENTS OTHER THAN MONEY. ARE THERE RESOURCES OR TOOLS OR OTHER THINGS THAT WE NEED TO KNOW ABOUT IN TERMS OF HUMAN CAPITAL THAT ARE NEEDED TO ADVANCE THESE AREAS. >> SO I ASK START WITH THE HEARING. -- FOR HEARING WE HAVE A LOT OF CHALLENGES. * TO BEGIN WITH -- SO WHEN WE -- THE INNER EAR IS NOT AN EASILY ACCESSIBLE ORGAN. WHEN I DO SURGERY -- THE INNER EAR WHEN WE GET TO THE POINT OF ACCESSING IT AND SURGERY FOR HEARING RESTORATION MANY PARTS OF IT ARE MISSING NOT JUST THE SENSORY CELLS BUT THE SUPPORTING CELLS SO THE DAMAGE IS NOT JUST ONE CELL TYPE THAT IS MISSING AND NOT A SINGLE CELL TYPE THAT YOU CAN TRANSPLANT AND PUT ANYWHERE. IT NEEDS TO BE IN A SPECIFIC LOCATION. WE STILL DON'T KNOW ALL OF THE REGULATORS AND NOT SUCCESSFULLY ENVY VOTE IN REGENERATING THE CELLS. THE REASON THEY LOSE THEIR SENSORY CELLS FOR SOME PEOPLE IT'S JUST A NOISE EXPOSURE OR TOXINS BUT THE VAST MAJORITY IT'S MUTATIONS. SO, ITS -- YOU GO BACK TO THE SAME PERSON AND YOU TRY TO REGENERATE THE CELLS. IF IT'S A CONGENITAL FORM OF HEARING LOSS THEN YOU NEED TO REPLACE OR CORRECT THE MUTATION. AND INDUCE THE NATIVE TISSUE TO DIVIDE AND REGENERATE. AND THEN THE LAST BARRIER IS THAT I'M SURE YOU'VE HEARD -- ABOUT SUCCESSFUL REGENERATION OF HAIR CELLS OR NOT REGENERATION BUT RESCUE OF HEARING LOSS IN DIFFERENT MOUSE MODELS. THESE ARE USUALLY MOUSE MODELS AND MUTATIONS AND HAIR CELL SPECIFIC GENES. AND THESE ARE SPECIFIC MUTATIONS BUT THE RESCUE OF THE HEARING ALMOST ALWAYS WITH THE EXCEPTION OF ONE GENE HAPPEN WHEN THE TREATMENT IS OFFERED WITHIN THE FIRST FIVE DAYS OF MOUSE LIFE. WHICH ISth EQUIVALENT OF FIRST TRIMESTER OF HUMAN PREGNANCY WHICH IS A TIME POINT WHERE A WOMAN WOULD NOT EVEN KNOW THAT SHE HAS AN EMBRYO WITH A PROBLEM. AND WHEN YOU TRY TO DO THE SAME THING LATER IT DOES NOT WORK. THE THOUGHT IS THAT THESE ARE EPIGENETIC CHANGES AND THE KEY FOR THAT IS ALMOST RELEASED. SO THERE IS SOMETHING TO LOOK FORWARD TO. THERE IS ONE MUTATION THE OTOF GENE WHERE THE HAIR CELLS ARE THERE AND THEY ARE RANKED GENERALLY INTACT BUT MISSING JUST THAT GENE PRODUCT. AND HEAR IS RESTORATION IS BEING SUCCESSFUL EVEN IN MATURE MICE THAT WERE INJECTED AND THAT IS -- THE FIRST GENE THAT HAS COMPETED ON BY THREE DIFFERENT FARM AS NOW TO GET TO THE CLINICAL TRIALS. >> MARTHA MAY I ASK RONNA A QUESTION. >> SURE. >> WHAT IS KNOWN ABOUT THE HALF LIFE OF THESE SENSORY HAIR CELLS IN HUMANS AND DO THEY REPLICATE AND TURN OVER AND IF THEY TURN OVER WHAT IS KNOWN ABOUT THE CELL THAT MAKES NEW ONES. >> THIS IS EXACTLY A PROBLEM. OUR HAIR CELLS ARE HOLD OLDER THAN US. THEY ARE POST MY NOTIC ALREADY IN THE -- TRIMESTER OF. IT'S THE SAME THAT WE HAD FROM BEFORE WE WERE BORN >> SO, IT'S SIMILAR TO THE PAN CRE YETIC BETA CELLS. THEY DIVIDE ABOUT SIX TIMES IN A PERSON'S LIFETIME SO THERE IS NO ADULT STEM-CELL FOR THE HAIR CELLS. >> THAT IS CORRECT. AND EARLY ON AND ALSO IN HUMAN -- IN MICE -- P34 THERE IS A THICKENING. IT'S CALLED THE GREATER ESPECIALLY THINKIAL -- AND YOU CAN -- INDUCE. *. THAT ENTIRE ARAB SOBS AND DISAPPEARS >> THAT IS REALLY VERY INTERESTING AND THANKS FOR BRINGING THAT UP DOUG BECAUSE THAT IS ANOTHER QUESTION THAT I HAD FOR THE PANEL. THE VALUE OF HUMAN EMBRYONIC STEM CELLS IN THE FIELD AND WHERE ARE WE GOING WITH THAT? BUT DOUG PLEASE I ASKED ABOUT CHALLENGES AND ROADBLOCKS SO SO MAYBE YOU WANT TO ADDRESS THAT FIRST BUT I WOULD LIKE TO MOVE ON TO THE HUMAN EMBRYONIC STEM-CELL QUESTION. BUT WHAT DO YOU THINK ARE THE CHALLENGES? >> IF I WOULD IDENTIFY ONE CHALLENGE IT WOULD BE THAT I'M CONVINCED THE COMMUNITY WILL FIND WAYS TO DIFFERENTIATE CELLS OF INTEREST. THEY CAN BE USED TO STUDY AND TREAT DISEASE. THE BIGGEST CHALLENGE IS TRANSPLANTATION AND DEALING WITH THE IMMUNE REJECTION. BUT I DO NOT BELIEVE WE'LL GET TO THE POINT WHERE YOU CAN COMMERCIALIZE PERSONALIZED MEDICINE. IT IS FAR TOO TEDIOUS AND EXPENSIVE. I COULD BE WRONG SO IF SOMEONE FIGURES OUT HOW TO MAKE AN IPSL FROM EVERY PATIENT AND DIFFERENTIATE IT INTO THE CELL OF INTEREST FOR EXAMPLE A SENSORY HAIR CELL AND IF THEY CAN DO THAT FOR A FEW HUNDRED THOUSANDS DOLLARS THEN I'M WRONG. AT PRESENT ESTIMATES WHAT I JUST SAID IS A $3 MILLION OR $4 MILLION PROJECT. BEFORE YOU GET TO TRIAL AND IMPLANTATION. TOLERANCE AND ALOE REJECTION ARE THE BIGGEST PROBLEMS. >> YEAH. I'M NOT DISAGREEING WITH YOU THERE. BUT YEAH -- I HAD THE OTHER QUESTION FROM OUR PREVIOUS DISCUSSION WHICH WAS THE USE OF HUMAN EMBRYONIC STEM CELLS AND RELEVANT TO HEARING LOSS AND DIABETES MANAGEMENT AND THERAPIES FOR THESE TWO CONDITIONS. WHERE ARE WE WITH THAT? >> I WOULD SAY I'M SURPRISED BY HOW LITTLE PROGRESS THERE HAS BEEN MADE IN THE FOLLOWING CONTEXT. I'M NOT GOOD AT DATES BUT LET'S SAY WE DO 25 YEARS AGO THAT HUMAN EMBRYONIC STEM CELLS WOULD BE POTENTIALLY USEFUL FOR REGENERATIVE MEDICINE. THE FACT IS THAT AT THIS MOMENT IN TIME THERE ARE FEW CLINICALLY COMPLIANT CELL LINES AVAILABLE AND THEY ARE HARD TO GET. THEY INVOLVE COMPLEX LICENSING AGREEMENTS. HARDLY ANY HAVE BEEN MADE. PLENTY OF RESEARCH LINES. I'M NOT TALKING ABOUT RESEARCH LINES BUT IF YOU WANT TO TRANSPLANT THINGS INTO PEOPLE AS YOU APPRECIATE THE CLINICALLY COMPLIANT LINE THE NIH HAS MADE, A FEW COMPANIES HAVE MADE SOME BUT IF YOU ASKED ME 25 YEARS AGO I WOULD GUESS THERE WOULD BE HUNDREDS AND THEY DON'T. THEY COME WITH RESTRICTIONS AND EACH LINE HAS A DIFFERENT BEHAVIOR WHEN YOU GO TO DIFFERENTIATE. SO YOU HAVE TO TRY A FEW LINES TO FIND THE ONE THAT IS GOING TO DO WHAT WANT. >> RONNA WHAT IS YOUR RESPONSE AND FOR BOTH OF YOU DO YOU HAVE ANY LAST PARTING WORDS OF WISDOM FOR THIS PANEL? >> ENVIOUS OF DOUG BECAUSE HIS CELLS CAN GROW IN -- AND HIS BIGGEST PROBLEM IS THE IMMUNE RESPONSE. WE REALLY LOOK AT STEM CELLS AS -- MODELS TO HELP US SCREEN FOR TOXICITY TO HELP US LEARN ABOUT REGULATORS OF CELL FATE -- TO HELP US EVEN IDENTIFY THE EPIGENETIC LANDSCAPE IF YOU WANT OF HUMAN HAIR CELLS. BECAUSE THE ONLY SOURCE FOR US FOR TISSUE IS FROM HUMAN IS -- ORGAN TRANSPLANT DONORS THAT WE CAN GET TISSUE FROM. AND THAT IS VERY, VERY DIFFICULT AND VERY -- AND THE OTHER OPTIONS -- -- TO BEGIN WITH THEIR CELLS ARE NOT NORMAL. SO WE USE STEM CELLS IN OUR FIELD AS INSTRUCTIVE CUES ALTHOUGH THERE ARE SOME GROUPS USING STEM CELLS POSSIBLY TO HELP REGENERATE KNE NEURONS >> DOUG ANY LAST WORDS OF WISDOM? >> I DON'T KNOW IF THEY ARE WORDS OF WISDOM. BUT MORE OF ENCOURAGEMENT. IF WE THINK ABOUT ALL OF THE DIFFERENT CELLS IN OUR BODY WHICH ARE DEGRADED WITH AGE LET ALONE DISEASE AND THE FACT THAT STEM CELLS CAN MAKE THOSE THE COMMUNITY SHOULD THINK MORE ABOUT TRYING TO DERIVE ONES OF INTERESTS. HOW MANY CELLS CAN BE MADE THAT ARE FUNCTIONAL. ONLY A HANDFUL. IT'S NOT A LONG LIST. SO MAYBE FOR THE YOUNG PEOPLE IN THE AUDIENCE I WOULD SAY PROBABLY THE MOST VALUABLE CELL YOU COULD MAKE WOULD BE A -- STEM-CELL. MAKE A UNIVERSAL DONOR -- THAT COULD GO INTO ANY PERSON AT ANY TIME. AS FAR AS I KNOW NO ONE HAS ACHIEVED THAT GOAL. NEITHER IN MOUSE LET ALONE HUMAN. SO, THAT IS A BIG CHALLENGE. GO OUT AND MAKE HUMAN -- STEM CELLS FROM A CLINICALLY COMPLIANT EMBRYONIC STEM-CELL LINE. THAT WILL HELP WITH ALL SORTS OF AGING AND DISEASES BEYOND BLOOD >> SO UNIVERSAL STEM-CELL HUMAN EMBRYONIC STEM-CELL DONOR. WE WOULD HAVE TO FIND A WAY TO IDENTIFY THE SUPER DONOR. 7 SO I'M SURE NOT EVERYONE IS QUALIFIED TO BE THAT UNIVERSAL DONOR. BUT FINDING WAYS TO IDENTIFY THE SUPER DONOR IDEA -- WOULD BE BENEFICIAL >> AND BEYOND THAT FIGURE OUT HOW TO MAKE IT INTO A HEMATOFATIC STEM-CELL AND THE BLOOD FIELD IS HARDLY WANTING FOR NUMBER OF INVESTIGATORS. IF YOU PUT BLOOD INTO THE PubMed YOU CANNOT DEAL WITH THE NUMBER BUT I DON'T THINK THAT ONE HAS MADE A -- STEM-CELL A FUNCTIONAL ONE FROM AN EMBRYONIC STEM-CELL >> WE'RE UP WITH TIME I THINK. WE'RE AT 5:10 AND I THINK WE'RE SUPPOSED DO A WRAP-UP AND ADJOURN. BUT I WOULD LIKE TO THANK YOU TWO. WONDERFUL PRESENTATIONS. SUCH EXPERT OPINIONS AND I APPRECIATE YOUR TIME AND EFFORTS HERE. >> THANK YOU. >> THANK YOU. -- ANYBODY ELSE? >> I WANT TO THAN CAN THE SPEAKERS AS WELL FOR THE FANTASTIC SESSION. VERY INFORMATIVE. AND VERY INVOLVING. -- EVOLVING. I'M HERE WITH MY PARTNERS IN CRIME. -- I WANT TO SHARE THE SCREEN QUICKLY AND -- TO HIGHLIGHT THE SUMMARY OF WHAT I THINK WE HEARD AND PLEASE CHIME IN AS YOU DEEM APPROPRIATE. IT'S ABOUT A FANTASTIC TWO DAYS INTO MANY TOPICS. WE TOUCHED ON A LOT OF DIFFERENT TOPICS. IT WAS VERY USEFUL TO SEE THE DIFFERENT FIELDS AND DIFFERENT LEVELS OF PRODUCT DEVELOPMENT. AS THIS FIELD IS EVOLVING AND BECOMING MATURE. IT IS IMPORTANT THAT WE LEARN FROM EACH OTHER. AND THERE IS CROSS POLLEN ASIAN. IT IS NONE * THE LESS A COMPLEX FIELD AND IT'S DIFFICULT TO TAKE THE COMPLEXITY. THROUGH THOSE DISCUSSIONS AND INTERACTIONS A LOT OF THE COMMONALITIES ARE EMERGING AND TRYING TO STREAMLINE SOMEHOW THE PROGRESS. WE LEARNED THAT IT TAKES A VILLAGE AND WE CANNOT DO THIS IF WE'RE IN ISOLATION. WE NEED TO SHARE AND PROTECT YOUR INTELLECTUAL PROPERTY BUT YOU -- IT IS NONETHELESS A MULTI-DISCIPLINARY APPROACH TO MAKE IT POSSIBLE. SO WE NEED THE SYNERGY. WHAT WE LEARNED ALSO IS THAT THERE WILL NOT BE A SILVER BULLET. WE NEED TO THINK AT THE GLOBAL LEVEL AND WE STILL NEED A LOT OF VERY SOLID BASIC SCIENCE IN ORDER TO MOVE AN IDEA FROM A CONCEPT TO A TREATMENT. AND WE LEARN IT FROM SUCCESS STORIES. ALSO -- THE IMPORTANCE OF BUILDING THE WORKFORCE. THIS IS ALL NEW AND WE NEED TO ALSO CONSIDER -- THAT WE NEED TO BRING PEOPLE FROM DIFFERENT DISCIPLINES -- AND ASKING THE QUESTIONS. AND -- -- AND GOING BACK TO THE -- APPROACH IN BUILDING THE COMMONALITIES THERE. WE LEARNED ALSO THAT THE REGULATORS -- ARE YOUR BEST FRIENDS IN THE SENSE THAT THEY ARE THERE FOR YOU AND THEY WILL TALK TO YOU ONCE YOU HAVE SOMETHING THAT IS CONCRETE. AND ALSO THAT BY WORKING TOGETHER WE CAN FIND THE RIGHT BIOMARKERS. -- TO RELEASE PRODUCTS -- SO THAT YOUR SOLUTION CAN MAKE IT TO THE -- AND THE -- DISSEMINATION IS NOT EASY. YOU'RE TRYING TO MAKE A SOLUTION FROM A SINGLE TAILORED FOR EACH PATIENT. ALSO SOMETHING THAT -- BE A BATTLE. THERE MIGHT BE OPPORTUNITIES AS THE TECHNOLOGY -- AND THAT WOULD INCREASE THE MOMENTUM AND BRING NEW SOLUTIONS FASTER AND MAYBE EXTRAPOLATE FROM ONE TO THE OTHER. THIS IS WHAT WE DO FOR A LIVING. THIS IS YOUR PROGRAM OFFICER TRYING TO HELP YOU FILL IN THE GAPS SO THAT YOU CAN WORK FROM ONE TO THE NEXT. THIS TYPE OF DISCUSSION IS REALLY HELPING US UNDERSTANDING WHAT ARE THE CHALLENGES THAT YOU AS A COMMUNITY ARE FACING AND HOW WE CAN HELP THE SCIENCE MOVE FORWARD AND -- FOCUSING ON BRING NEW SOLUTIONS FOR OUR PATIENTS. THIS IS WHAT -- THAT IS WHY WE GO TO THE GYM EARLY. MAYBE WE'LL GET HERE TO -- --- ---THIS IS A VERY OLD CARTOON. IT'S BEEN AROUND FOREVER. IT'S A LONG WAY BUT WE ARE CLOSER TO HAVING THE POSSIBILITY. SO -- BY ALL MEANS I CAN BE SUPER EXHAUSTIVE OF THESE. I WANT TO TELL YOU THIS IS JUST THE STARTING POINT. WE WANT THIS TO CONTINUE. I NEED TO THANK THE SPEAKERS. THEY DID A FANTASTIC JOB IN PROVIDING THEIR PERSPECTIVES IN THEIR OWN FIELD BUT ALSO CONSIDERATIONS THAT CAN BE APPLIED TO A LOT OF FIELDS. -- -- A FANTASTIC JOB IN STIMULATING QUESTIONS AND COORDINATING THE DISCUSSION. I WANT TO THANK THE PLANNING COMMITTEE WHO HELPED PLAN THE TOPICS AND THE SPEAKERS TO HAVE A SOUND MEETING. THAT I THINK WAS VERY WELL ORGANIZED. THE NICHD LEADERSHIP -- -- AGAIN THE CHARGE OF -- THIS MEETING -- -- ----- ALSO WANT TO THANKING THE COMMUNICATION TEAM AND THE LOGISTIC TEAM AND CHRISTINA -- ALICIA -- ALEX SIMPSON. KATHERINE TRAINER AND -- -- -- I ALSO WANT TO THANK THE AUDIENCE. THAT HAS BEEN WITH US FOR QUITE SOME TIME AND I WANT TO MENTION THAT EVERYBODY IS AWARE THAT THE MEETING IS BEING RECORDED. IT WILL BE AVAILABLE ON THE NIH VIDEO CAST WEBSITE. UNDER CONFERENCES AND WE WILL HAVE IT ON THE NICHD WEB PAGE -- SO THAT PEOPLE WILL BE ABLE TO GO BACK AND SEE THOSE. BUT AGAIN THIS IS A STARTING POINT. IT'S NOT THE END. WE WANT YOU FOR CONTINUE TO BE ENGAGED. EVERYBODY CAN GO AND POST THEIR IDEAS AND COMMENTS. ENGAGE WITH EACH OTHER'S IDEAS AND COMMENTS. AND REALLY IT'S A WAY FOR YOU TO INFORM THE NIH OF -- -- -- IDEAS OF WHAT YOU THINK THE FIELD SHOULD GO IN AND THE NEEDS -- OF YOUR OWN FIELD IN ORDER TO MAKE IT TO THE NEXT STEP. AND WE REALLY VALUE YOUR FEEDBACK. IT IS VERY USEFUL TO PREPARE OUR FUTURE INITIATIVES AND OUR STRATEGIES TO HELP DO OUR JOB. FILL IN THE GAPS AS I MENTIONED. SO PLEASE ENGAGE IN THE CAMPAIGN. THERE IS NO SILLY QUESTION OR SILLY COMMENT AND WE WILL LOOK INTO EVERYTHING AND IF YOU HAVE ANY SPECIFIC QUESTIONS YOU CAN REACH OUT TO US ALSO AND BRING YOUR OWN IDEAS. SO I WANT TO THANK EVERYBODY. I THINK WE HAVE -- A -- -- WE NEED SOME TIME TO PROCESS AND UNPACK ALL OF THE INFORMATION THAT WAS HERE. A BIG SHOUT OUT TO OUR SPEAKERS. AND THANK YOU. THANK YOU FOR YOUR REPORTING.