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ESC-Derived Cells For Restoration of Tissues & OrgansRobert Lanza, MDVP Research & Scientific DevelopmentAdvanced Cell Tec...
Alzheimer’sDwarfismParkinson’sStrokesEpilepsyHemophiliaKidney failureChronic painCancerInfertilityBurnsAIDSMuscular dystro...
Anatomy & Function of RPE• Immune barrier• Absorption of stray light• Vit A metabolism & transport• Phagocytosis of shedph...
Diseases Associated with RPE dysfunction• Age-related macular degeneration• Retinitis pigmentosa• Stargardts disease• Best...
Transplantation of RPE in HumansAssociated problemsSources of RPE cells* autologous tissue* cell lines* donor tissue (adul...
Advantages of ECS-derived Tissues for RegenerativeMedicine• Unlimited supply• Can be derived under GMP conditions pathogen...
[All hES cell lines studied reproducibly generated RPE linesthat could be passaged, characterized, and expanded]•WiCell hE...
Stages of RPE isolation from spontaneously differentiating hES cells35 mm plate one of the clustersone of the clusters cel...
x400x200hES-RPE express RPE markers (bestrophin &CRALBP)-- 32-- 46-- 78CRALBPMwa b cbestrophinbestrophinCRALBPImmunostaini...
PEDF RPE65RT-PCR1 2 1 21 – fetal RPE2 – hES-RPEhES-RPE express RPE65 and PEDF
DBX15,200x7000Phagocytosis of latex beads (electron microscopy)Latex beadspigment
hES-RPE vs. its in vivo counterpartRPE hES-RPEcobblestone, pigmentedtransdifferentiation-differentiationphagocytosismolecu...
Gene expression profiling of hES-RPE vs. their in vivocounterpart
RPE transplantation into subretinal space of RCS rats(in collaboration with Raymond Lund, University of Utah)RCS rats natu...
1 2 3 1 2 3 1 2 3 1 2 3 1 2 3 1 2 3RPE65bestrophinCRALBPPEDFPax6GAPDHH9 RPE used for transplantation in RCS rats
conehES-RPEERG at P60Amplitude(uV)40a-hES-RPE a-sham b-hES-RPE b-sham coneb-sham1800206080120140160100hES-RPE transplantat...
Summary• hES-RPE is similar to its in vivo counterpart by multiple parameters(morphology, behavior, phagocytosis, molecula...
Hemangioblasts derived from hESC
Hemangioblasts Generated from Human ES Cells40X 10X
• Established an efficient method to generatehemangioblasts from multiple hESC lines• Cells can be easily & reproducibly s...
Generation of ES cells using parthenogenesis
WBC colony from cloned stem cells
•Cardiovascular disease costs the US $329 billion annually
10203040506070809010019881989199019911992199319941995199619971998199920002001YearNumberofPatients(inthousands)Waiting List...
End-stage renal disease will cost US $1 trillion during the comingdecade
6th Annual Stem Cells & Regenerative Medicine Meeting Conference Presentation
6th Annual Stem Cells & Regenerative Medicine Meeting Conference Presentation
6th Annual Stem Cells & Regenerative Medicine Meeting Conference Presentation
6th Annual Stem Cells & Regenerative Medicine Meeting Conference Presentation
6th Annual Stem Cells & Regenerative Medicine Meeting Conference Presentation
6th Annual Stem Cells & Regenerative Medicine Meeting Conference Presentation
6th Annual Stem Cells & Regenerative Medicine Meeting Conference Presentation
6th Annual Stem Cells & Regenerative Medicine Meeting Conference Presentation
6th Annual Stem Cells & Regenerative Medicine Meeting Conference Presentation
6th Annual Stem Cells & Regenerative Medicine Meeting Conference Presentation
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6th Annual Stem Cells & Regenerative Medicine Meeting Conference Presentation

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"ESC-Derived Cells for Restoration of Tissues & Organs"

10/16/2006

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6th Annual Stem Cells & Regenerative Medicine Meeting Conference Presentation

  1. 1. ESC-Derived Cells For Restoration of Tissues & OrgansRobert Lanza, MDVP Research & Scientific DevelopmentAdvanced Cell Technologyand Adjunct ProfessorWake Forest University School of Medicine
  2. 2. Alzheimer’sDwarfismParkinson’sStrokesEpilepsyHemophiliaKidney failureChronic painCancerInfertilityBurnsAIDSMuscular dystrophyALSAffective disordersMacular degenerationHypoparathyroidismHeart diseaseLiver failureEnzymatic defectsDiabetesOsteoarthritisMultiple sclerosisHuntington’sHypocholesterolemiaRheumatoidarthritisAtherosclerosisUlcersSpinal cordinjuries
  3. 3. Anatomy & Function of RPE• Immune barrier• Absorption of stray light• Vit A metabolism & transport• Phagocytosis of shedphotoreceptor segmentsFUNCTIONS OF RPE
  4. 4. Diseases Associated with RPE dysfunction• Age-related macular degeneration• Retinitis pigmentosa• Stargardts disease• Bests vitelliform macular dystrophy• Lebers congenital amaurosis
  5. 5. Transplantation of RPE in HumansAssociated problemsSources of RPE cells* autologous tissue* cell lines* donor tissue (adult, fetal) safety ethical Batch-to-batchvariationmay have impaired function limited supplyPotential tumorigenicity
  6. 6. Advantages of ECS-derived Tissues for RegenerativeMedicine• Unlimited supply• Can be derived under GMP conditions pathogen-free• Can be produced with minimal batch to batchvariation• Can be thoroughly characterized to ensure optimalperformance
  7. 7. [All hES cell lines studied reproducibly generated RPE linesthat could be passaged, characterized, and expanded]•WiCell hES cell lines (23 RPE lines generated)WA01 WA09WA07•Harvard hES cell lines (22 RPE lines generated)HUES1 HUES6HUES2 HUES7HUES3 HUES8HUES5 HUES10•ACT hES cell lines (25 RPE lines generated)MA01 MA03 MAJ1MA04 MA09MA14 MA40RPE can be generated from hES cells
  8. 8. Stages of RPE isolation from spontaneously differentiating hES cells35 mm plate one of the clustersone of the clusters cell suspension at plating4 daysx100x200x2007 daysx200Passage 1 -- 25 daysPassage 1 -- 25 daysx200x0.75
  9. 9. x400x200hES-RPE express RPE markers (bestrophin &CRALBP)-- 32-- 46-- 78CRALBPMwa b cbestrophinbestrophinCRALBPImmunostainingWestern blot
  10. 10. PEDF RPE65RT-PCR1 2 1 21 – fetal RPE2 – hES-RPEhES-RPE express RPE65 and PEDF
  11. 11. DBX15,200x7000Phagocytosis of latex beads (electron microscopy)Latex beadspigment
  12. 12. hES-RPE vs. its in vivo counterpartRPE hES-RPEcobblestone, pigmentedtransdifferentiation-differentiationphagocytosismolecular markersRPE65CRALBPbestrophinPEDFMERTK
  13. 13. Gene expression profiling of hES-RPE vs. their in vivocounterpart
  14. 14. RPE transplantation into subretinal space of RCS rats(in collaboration with Raymond Lund, University of Utah)RCS rats naturally become blind in several weeksdue to RPE degeneration and photoreceptor deathStudy designcell line RPE (H9)Control: culture mediumTests:head tracking (behavior)electroretinogram (ERG)histologyIn vitro assessment:molecular markers of RPEmorphology and behavior
  15. 15. 1 2 3 1 2 3 1 2 3 1 2 3 1 2 3 1 2 3RPE65bestrophinCRALBPPEDFPax6GAPDHH9 RPE used for transplantation in RCS rats
  16. 16. conehES-RPEERG at P60Amplitude(uV)40a-hES-RPE a-sham b-hES-RPE b-sham coneb-sham1800206080120140160100hES-RPE transplantation into subretinal space of RCS ratsOptomotor at P100hES-RPE Sham Untreated0.50.30.40.200.1Relativeacuity(c/d)
  17. 17. Summary• hES-RPE is similar to its in vivo counterpart by multiple parameters(morphology, behavior, phagocytosis, molecular markers)• hES-RPE can be reproducibly generated from hES cells• hES-RPE attenuates photoreceptor loss in animal model of retinal degenerationhES-RPE advantages• Minimize batch-to-batch variation• Can be derived under GMP conditions• Can be produced from feeder-free hES cells• Can be easily generated in large quantities(for pathogen & safety assessment, and pre-clinical & clinical studies)
  18. 18. Hemangioblasts derived from hESC
  19. 19. Hemangioblasts Generated from Human ES Cells40X 10X
  20. 20. • Established an efficient method to generatehemangioblasts from multiple hESC lines• Cells can be easily & reproducibly scaled up• Hemangioblasts derived from hESC formfunctional hematopoietic & endothelial cellsin vitro• Rapidly repair damaged vasculature in vivo(ischemic limbs, retina, and myocardium)Summary
  21. 21. Generation of ES cells using parthenogenesis
  22. 22. WBC colony from cloned stem cells
  23. 23. •Cardiovascular disease costs the US $329 billion annually
  24. 24. 10203040506070809010019881989199019911992199319941995199619971998199920002001YearNumberofPatients(inthousands)Waiting ListOrgans Transplanted
  25. 25. End-stage renal disease will cost US $1 trillion during the comingdecade

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