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Exploring the Potential of Stem Cells in Regenerative Medicine

Discover the role of stem cells in embryo development and adult body, their self-renewal capacity, and differentiation characteristics. Learn about stem cell niches, transcriptional networks, and various modes of neurogenesis. Delve into the groundbreaking work of scientists like John Gurdon and Shinya Yamanaka in stem cell research.

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Exploring the Potential of Stem Cells in Regenerative Medicine

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  1. Stem cell and regenerative medicine 5-9-2016

  2. Why we need stem cells in our body? In embryo development: to increase cell number and to differentiate into different cell types. In adult body: to continuously supply short lived somatic cells and to replace damaged cells

  3. Stem Cells • Stem cells have two defining attributes: • The capacity for self-renewal. • The ability to differentiate into many different cell types.

  4. Stem Cells • Niche of stem cells

  5. Adult stem cells in tissue are thought to reside in niches

  6. Patterns of stem-cell differentiation.

  7. Transcriptional network regulating pluripotency of ES cells.

  8. Embryonic stem (ES) cells can be maintained in culture and can form differentiated cell types.

  9. Embryonic stem cells in the dish

  10. Intestinal stem cells:ｈｏｗ　ｔｏ　ｉｄｅｎ.

  11. Regeneration of the intestinal epithelium from stem cells can be demonstrated in pulse-chase experiments.

  12. Three modes of neurogenesis during cortical development. RG in cortex generate neurons

  13. Neural stem cell niche in the adult brain.

  14. Formation of blood cells from hematopoietic stem cells in the bone marrow.

  15. The original question: Do all cells in the body have the same sets of genes?

  16. Two alternative hypothesis: or

  17. PNAS 1952 38 (5) 455-463

  18. 是否發育晚期的細胞核已無法回復全能發育的潛力？是否發育晚期的細胞核已無法回復全能發育的潛力？ 丟失了基因還是不可逆的修飾？

  19. John Gurdon Kt DPhil DSc FRS Distinguished Group Leader in the Wellcome Trust/CRUK Gurdon Institute in Cambridge

  20. 戈登1933年出生。 • 他中學唸的是1440年創校的伊頓公學 (Eton College)。 • 在校成績生物科在250位同級生中名列最後。 • 導師給他的評語是：我想戈登希望未來成為一位科學家，但從他目前的表現看來，這似乎是個不可能的任務！ • 進牛津大學專攻古典文學，隨後轉到動物系 • 1961年得到動物學的博士學位。 • Studies on nucleocytoplasmic relationships during differentiation in vertebrates . • 美國加卅理工學院，作了有關細菌遺傳學的博士後研究。 • 1962年回到英國牛津大學動物系任教，1971轉到劍橋大學任教迄今。

  21. Why should Gurdon repeat this experiment?

  22. Two technical innovations: 1,用Zenopus的卵 (big size!) 2, 用UV來破壞卵中的染色體和去除外層保護的膠質

  23. Late stage embryonic cell nuclei still can develop an viable frog! How about a fully differentiate cell nuclei?

  24. 完全分化的體細胞仍然擁有完整的遺傳資訊！ Gurdon, J.B. & Uehlinger, V. Nature 210, 1240–1241 (1966)

  25. What is next?

  26. Is somatic cell nuclear transfer (SCNT) also true for mammalian animal?

  27. Dolly, 1997

  28. Is Dolly a scientific breakthrough?

  29. What are signals in the cytoplasma of egg to trigger reprogramming of silenced genetic program in somatic cell nuclei?

  30. Could we identify cytosolic factors which can convert a somatic cell nuclei into a embryonic stem cell with totipotent as a fertilized egg ？ A mission impossible!

  31. Shinya Yamanaka • 1962年出生. • 1987年在 Kobe University得到MD. • 1993在Osaka City University 得到Ph.D. • 1993-96 在UCSF作博士後研究 • 1996 – 99Osaka City University 助理教授. • 1999–2003Nara Institute of Science and Technology副教授. • 2004 -10 Institute for Frontier Medical Sciences教授 • 現任京都大學教授和 iPS Cell研究與應用中心主任

  32. Identification of ES cell associated transcriptsby digital differential display To compare expressed sequence (EST) libraries from mouse ES cells (3 libraries; 33,077 clones) and those from various somatic tissues (103 libraries; 1,040,493 clones) and to identify genes overrepresented in ES cell-derived libraries. http://www.ncbi.nlm.nih.gov/UniGene

  33. Transgenic fibroblasts with a knock-in gene at the Fbx15 locus. The knock-in was βgeo: a fusion of LacZ gene and neomycin-R gene Normally, Fbx15 is highly expressed in ES cells, but not expressed in fibroblasts

  34. 24 genes were selected as candidates to induce pluripotency in somatic cells, For b-catenin, c-Myc, and Stat3, they used active forms, S33Y-b-catenin, T58A-c-Myc and Stat3-C, respectively. Because of the reported negative effect of Grb2 on pluripotency, they also included its dominant-negative mutant Grb2DSH2 as 1 of the 24 candidates.

  35. Are they really pluripotent stem cells ? Teratoma formation and germ line differentiation in vivo Embryoid body formation and differentiation in vitro

  36. Induction of Pluripotent Stem Cellsfrom Mouse Embryonic and AdultFibroblast Cultures by Defined Factors Kazutoshi Takahashiand Shinya Yamanaka Cell 126, 663–676; 2006

  37. Elite vs Stochastic model

  38. Reprogramming occurs in two phases

  39. Stoichiometry of factors is important for reprogramming

  40. Two major application of iPS Personalized drug screening Personalized cell therapy

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