2. MARIE CSETE MD, PhD CHIEF SCIENTIFIC OFFICER mcsete@cirm.ca.gov

3. CANCER SURVIVAL Early Development DRUG DEVELOPMENT TOXICICOLOGY TRANSPLANTATION DEGENERATIVE DISEASES TRAUMA: Spinal cord Burns Head injury AGING TISSUE ENGINEERING

4. ALL STEM CELLS CAN EITHER -SELF-RENEW and/or -GENERATE SEVERAL KINDS OF CELL TYPES

5. SELF-RENEWAL Stem X X X X

6. BLOOD STEM CELLS ARE PLURIPOTENT

7. “-POTENCY” Multipotent: A few Pluripotent: A lot Totipotent: Every kind of cell including cells of the body (somatic) and cells of the germline (eggs, sperm)

8. TOTIPOTENT ONLY EMBRYONIC STEM CELLS ARE CONSIDERED TOTIPOTENT BUT REALLY THEY ARE NOT TOTIPOTENT BECAUSE OF THE ISSUES OF COMPLEX ORGANIZATION ONLY THE FERTILIZED EGG IS REALLY TOTIPOTENT

9. WHERE DO YOU GET THEM? ADULT STEM CELLS -ANY STAGE OF EMBRYO -ANY STAGE OF ADULTHOOD EMBRYONIC STEM CELLS -BLASTOCYST EMBRYOS -SCNT: Somatic cell nuclear transfer -PARTHENOTES -iPS: Induced pluripotent cells

10. University of Wisconsin website

11. University of Wisconsin, 2001.

14. . University of Wisconsin, 2001. Embryonic stem cells are derived from the inner cell mass of the blastocyst stage embryo

15. University of Wisconsin, 2001. In the lab, trophoblast is removed and the embryonic stem cells singly selected from the inner cell mass

16. University of Wisconsin, 2001. ES cell cultivation in the laboratory: easier with mouse than human cells

17. University of Wisconsin, 2001. Using a variety of tools, in theory any cell type can be made

19. HUMAN EMBRYONIC STEM CELLS.

20. Where they come from Potency Proliferative senesence (source material abundance) EMBRYONIC VS. ADULT STEM CELLS Important differences :

21. 1997—DOLLY and THE ERA OF CLONED ANIMALS

22. +

23. ADAPTED FROM WWW.NIH.GOV/NEWS/STEMCELL/FIG4B.GIF Blastocyst Inner Cell Mass (Pluripotent) Egg Cell (Remove Nucleus) (Fusion) Somatic Cell Nuclear Transfer (Extract Inner Cell Mass) Cultured Pluripotent Embryonic Stem Cells Somatic Cell Nuclear Transfer: IS NOT HUMAN CLONING Somatic Cell nucleus (any cell in the body other than an egg or germ cell) (Stimulate Cell Division Process) ADAPTED FROM WWW.NIH.GOV/NEWS/STEMCELL/FIG3.GIF

24. Patient advocacy for SCNT No immunologic barrier with cells generated from recipient Screening drugs for many common diseases using clones from families with inherited diseases First glimpse into early development Does not require use of embryos

27. Use for parthenogenetic lines Genetic homozygosity Bank cells with specific MHC proteins Other uses? Engineer cells for HIV resistance  make HSC for marrow transplantation SCNT AND PARTHENOGENESIS REQUIRE AN EGG Human egg donation is not trivial

28. All 3 germ layers represented -ectoderm -mesoderm -endoderm Functional diff cell types

29. How/why would this work? Huge interconnectedness of protein networks Changes silencing status of whole genome: These factor awaken silenced genes all over the chromosomes Ultimate test of pluripotency: Make a whole organism

30. iPS lines used to make mice Reported simultaneously by 3 groups -Jaenisch (MIT) -Hochedlinger (Harvard)* -Yamanaka (Japan) *20% of the 121 offspring developed tumors -viral vectors? -not from vectors?

31. Hard problems What are network requirements for totipotency? Still left with little control over differentiation –Transcription and environment –Functional integration into tissue ES vs. cancer: Many overlapping traits Scale-up/manufacture: unsolved

32. Making ES cells easier than controlling them: MUSCLE

33. ? muscle fat Positive and negative signals redox reduceoxidize

34. CONTROL 2. GUTSKINCARDIACMUSCLESKELETALMUSCLEPANCREATICBETACELLS HEPATOCYTESNEURONSSCHWANNCELLSASTROCYTESSMOOTHMUSCLE BONEPLATELETSREDCELLSWHITECELLSKUPFFERCELLSTENDON

35. Complex development Recapitulate the uterine environment and signals –Engineering interface critical Recapitulate the developmental course ES  definitive endoderm  foregut endoderm  pancreatic endoderm  insulin producing mature beta cells (Dr. Melissa Carpenter, NOVOCELL)

36. Growth cues to imitiate complex organization of embryos

37. Critical issues for translation OLIGODENDROCYTES ASTROCYTES ? Transplant progenitors  risk of astrocyte scar Transplant mature oligodendrocytes  they don’t survive transplant

38. Tumors and stem cells Common traits: Undifferentiated, pluripotency, highly proliferative, self- renewing, migration, common markers Teratomas Permissive environments Cancer stem cells are those likely to escape treatment Ultimately understanding stem cells will lead to enormous gains in cancer biology Chemotx wipes out stem cells?

39. Inner core: Slowly dividing neuroepithealial cells— Roy et al, Nature Med 2006