Koç Medicine Phase i Lecture Dec 24, 2010

1. CURRENT PERSPECTIVES ON STEM CELL BIOLOGY AND FUTURE IMPLICATIONS Emin Kansu,MD,FACP [email_address] Hacettepe University Institute of Oncology

2. 1953

3. NATURE February 15,2001 HUMAN GENOME MAP

4. CLONING TECHNOLOGY

5. DOLLY First Cloned Animal- 19*****

9. HIROSHIMA ve NAGASAKI August 1945 A-BOMB

10. BALB/c BONE MARROW CELLS MOUSE SPLEEN IRRADIATED MOUSE BONE MARROW BALB/c BALB/c IRRADIATION WILL EMPTY BONE MARROW SPACE AND RECONSTITUTION CAN BE ACHIEVED BY NEW STEM CELLS CFU-S INFUSION OF IRRADIATION

12. FROM EMBRYONIC STEM CELLS TO A NEWBORN Human Embryonic Stem Cells (hESC)

16. STEM CELL STEM CELL SYMMETRIC STEM CELL DIVISION STEM CELL

17. STEM CELL DIFFERENTIATED CELL STEM CELL ASYMMETRIC STEM CELL DIVISION

18. STEM CELL SELF-RENEWAL STEM CELL DIFFERENTIATION DIFFERENTIATED CELL (e.g.Myocyte, Neuron)

19. STEM CELLS I.Weissmann’05 DIFFERENTIATION SELF-RENEWAL MITOSIS APOPTOSIS G 1 Damar MIGRATION MOBILIZATION > 30.000/DAY (1-5 mins))

20. SPERM + OVUM DEVELOPMENT OF EMBRYO AND NEWBORN FERTILIZATION ESC BLASTOCYTE

21. EMBRYONIC STEM CELLS ECTODERM MESODERM ENDODERM TISSUE SPECIFIC ADULT STEM CELLS DIFFERENTIATED MATURE CELLS

29. HOW THEY ARE TRAINED FOR SELF-RENEWAL AND PLURIPOTENCY ?

30. SITES OF HEMATOPOIETIC STEM CELLS YOLK SAC AGM FETAL LIVER BONE MARROW PRIMITIVE ERYTROPOEISIS PLACENTA

31. HEMATOPOIETIC STEM CELLS PLURIPOTENT STEM CELL MESODERM Hemangioblast HEMO ENDODERM HSC EMBRYO 7.5 DAYS 10.5 12.5 15.5 BIRTH PRIMITIVE STREAK YOLK SAC PLACENTA FETAL LIVER BONE MARROW MICROENVIRONMENT DEVELOPMENT OF HEMATOPOIETIC STEM CELL

32. G O RESTING “ Dormant” STEM CELL DIFFERENTIATION SENESCENCE SPECIFICATION CELL CYCLE APOPTOSIS G 1 G 0 M HEMATOPOIETIC STEM CELL POOL

33. DEVELOPMENT OF STEM CELL “ NICHE” YOLK SAC PLACENTA FETAL LIVER BONE MARROW NICHE : ENDOTHELIAL CELLS OSTEOBLASTS Trabecular Bone

34. STEM CELLS PROGENITORS DIFFERENTIATED MATURE CELLS

35. STEM CELLS PROGENITORS DIFFERENTIATED CELLS Decline in Replication Potential Increasing Degree of Differentiation STEM CELL DIFFERENTIATION

36. SELF-RENEWAL Self-Renewal : no Mitotic activity : ++ *** Self-Renewal :no Mitosis : no SC PC DC

37. HEMATOPOIETIC STEM CELLS (HSC)

38. HEMATOPOIETIC STEM CELLS RED CELLS MONOCYTES PLATELETS LYMPHOCYTES EOSINOPHILS NEUTROPHILS BASOPHILS

39. THYMUS BONE MARROW HEMATOPOETIC STEM CELL

40. BONE MARROW UNIVERSITY (State)

41. THYMUS UNIVERSITY PROLIFERATION DELETION SELECTION T-LYMPHOCYTE FACULTY

42. MICROANATOMY OF BONE MARROW

43. BONE MARROW UNIVERSITY – INTERNAL STRUCTURES Central Sinus

44. BONE MARROW : “ UNIVERSITY CAMPUS” SINUS Adventitial cell My ERYT ( Mesodermal Stem Cell) CELLS

45. Stem Cell Education Site = NICHE 1978

46. HEMATOPOIETIC STEM CELLS “ NICHE” “ Education Sites for Stem Cells”

47. BONE MARROW “NICHE ” SINUS Adventitial cell My ERYT ( Mesodermal Stem Cell) CELLS NICHE

48. “ EDUCATION ” OF HEMATOPOIETIC STEM CELLS AT “NICHE” SITE “ NICHE” MHC HEMATOPOIETIC STEM CELL DC CD4+ T- Cells CD8+ T- CElls NK Cells NKT Cells TReg

49. STEM CELL AT THE “ NICHE ” STEM CELL Myeloblast BONE MARROW STROMA “ NICHE” MYELOID CELLS

50. SELF-RENEWAL CAPACITY LT-HSC ST-HSC MPP NK+ Cell T Cell B Cell GIMP MEP PMN MEG PLTS RBC CLP CMP

51. HEMATOPOIETIC STEM CELLS-MOUSE LT-HSC ST-HSC Progenitors I.Weissman’05 Common Lymphoid Common Myeloid

53. HEMATOPOIETIC STEM CELLS RED CELLS MONOCYTES PLATELETS LYMPHOCYTES EOSINOPHILS NEUTROPHILS BASOPHILS

55. OCT 4 NANOG SOX 2 DEVELOPMENTAL REGULATORS PROGRAMMING DIFFERENTIATION

57. PERIPHERAL BLOOD LYMPHOCYTES Stem Cell ?

59. STEM CELL MARKERS : MOUSE vs HUMAN c-kit + Thy1 low sca-1 + c-kit + Thy1 low CD34 + NEGATIVE B 220 Mac-1 Gr-1 CD3 CD4 CD8 Ter 119 NEGATIVE CD38 CD10 CD14 CD15 CD16 CD19 CD20 I.Weissman

60. STEM CELLS……

64. adapted from : Korbling & Estrov, NEJM 2003 ADULT STEM CELLS

66. HEMATOPOIETIC STEM CELLS RED CELLS MONOCYTES PLATELETS LYMPHOCYTES EOSINOPHILS NEUTROPHILS BASOPHILS

67. ALLOGENEIC STEM CELL TRANSPLANTATION RECIPIENT HEALTHY PERSON HEMATOPOIETIC STEM CELLS

68. ANNUAL NUMBERS OF BLOOD AND MARROW TRANSPLANTS WORLDWIDE 1970-2000 MDM01_21.ppt NUMBER OF TRANSPLANTS YEAR 1970 1975 1980 1985 1990 1995 Autologous Allogeneic 2000 0 5,000 10,000 15,000 20,000 25,000 30,000 35,000 40,000

69. E.DONNALL THOMAS, M.D. Emeritus Professor of Medicine University of Washington Fred Hutchinson Cancer Research Center - Seattle, USA Nobel Laureate for his contributions to Hematopoietic Stem Cell Transplantation in hematologic disorders 1990 1990

71. Morphology of Mesodermal Progenitor Cells (MSC) (CD 45- GlyA-)

72. MESENCHYMAL STEM CELLS BONE CARTILAGE MUSCLE TENDON LIGAMENT CONNECTIVE TISSUE Chondroblasts Collagen II SM Myoblast Sm Myosin CaPo4 Osteoblasts

74. FROM EMBRYONIC STEM CELLS TO A NEWBORN Human Embryonic Stem Cells (hESC)

75. Sir MARTIN EVANS University of Cardiff U.K.

76. MOUSE EMBRYONIC STEM CELLS Martin Evans’71 & 81

77. 2007 Physiology and Medicine Oliver Smithies Univ North Carolina U.S.A. Mario Capecchi Univ of Utah U.S.A. Sir Martin Evans Cardiff Univ U.K. “ Contributions to Embryonic Stem Cell Biology and Development of Knock-Out Mouse Model to Understand Pathobiology of Diseases”

79. SPERM + OVUM DEVELOPMENT OF EMBRYO AND NEWBORN BABY FERTILIZATION

80. IVF TÜPLERİNDEN EMBRYONİK KÖK HÜCRE ELDESİ IVF In Vitro Fertilization Thomson J & Yu SCIENCE’1998 :1998

81. ISOLATION OF EMBRYONIC STEM CELLS FROM INNER CELL MASS OF A BLASTOCYTE

82. EMBRYONIC STEM CELL COLONY

83. In-Vitro Differentiation of hESCs Embryonic Stem Cells Neurons-Tubulin + Cardiomyocytes Cardiac troponin+ Endoderm-AFP +

84. IN VIVO DIFFERENTIATION OF hESCs SCID/beige mice hESCs

87. EMBRYONIC STEM CELL RESEARCH- VATICAN’S VIEWS SHOULD THE RESEARCHERS USE HUMAN EMBRYONIC CELLS ? ANSWER : NO IS IT ETHICAL TO CLONE EMBRYONIC CELLS AND THEN DESTROY THEM FOR RESEARCH ? ANSWER : NO IS IT ETHICAL TO USE HUMAN ESCs ? ANSWER : NO

88. Shinya Yamanaka,M.D. “ Turning Back the Cellular Clock...” Takashi K, et al,Cell 2006

90. SKIN BIOPSY FROM TAIL OF MOUSE CELLS ARE SEPERATED SKIN FIBROBLASTS ARE OBTAINED NUCLEUS IS REMOVED Takashi & Yamanaka ‘2006 MOUSE OOCYTE NUCLEAR TRANSFER Enucleated Oocyte

91. “ iPS CELLS” Embryonic –like Cells GFP+ FROM SKIN to iPS CELLS Cell Cultures OCT-4 SOX-2 Kit-4 c-myc

92. November 20, 2007 Re-Programming Adult Stem Cells (Skin Cells) to Embryonic like cells Without Cloning and without Making Embryos Inducible Pluripotent Stem Cells (iPS)

93. I nducible P luripotent S tem Cells ( iPS Cells ) S.Yamanaka J.Thompson Re-Programming of Somatic Cells Rudolph Jaenisch

94. Breakthrough of the Year REPROGRAMMING CELLS Science 19 December 2008

95. STEM CELL RESEARCH

96. BENCH  BED-SIDE

97. THE STEM CELL DREAM

98. ARTIFICIAL BLADDER …April 2006 Wake Forest University Winston –Salem , California HARVARD MEDICAL SCHOOL BOSTON STEM CELLS AND REGENERATIVE MEDICINE

99. FACE TRANSPLANTATION 2005

100. CARDIAC REPAIR BY STEM CELLS

101. Growing Human MSCs on Nanofibril Scaffolds Chen FH,et al.Nature 2006

103. STEM CELLS WHICH CELLS ? NUMBER OF CELLS ? AT WHICH STAGE OF DISEASE ? TO INDUCE A REMISSION ? OR FOR CURE ? OR FOR PALLIATION ?

105. ARE WE READY TO USE EMBRYONIC STEM CELLS ? IN-VITRO YES IN-VIVO Animals Yes IN-VIVO FOR HUMANS ? NO ! NO!

107. MEDICINE The Whole World Is Watching Hope—and anxiety—run high as the first clinical trial of embryonic-stem-cell therapy begins this summer. President Barack Obama’s executive order lifted restrictions on Embryonic Stem Cell Research NEWSWEEK March 30 , 2009

108. FUTURE PROSPECTS OF STEM CELL RESEARCH TO UNDERSTAND THE DEVELOPMENTAL BIOLOGY (Endoderm-Mesoderm-Ektoderm) FOR SCIENTIFIC BASIS OF CELLULAR THERAPIES TO UNDERSTAND PATHOPYSIOLOGY OF HUMAN DISEASES DRUG DISCOVERY AND TOXICOLOGY TESTING

109. More Basic Research is Needed… Too Many Hopes.... Future of Stem Cells The ideas are Great !

112. CD4+ T-CELLS CD8+ T- CELLS NK CELLS NKT CELLS Treg CELLS HEMATOPOIETIC STEM CELL HOMING AND EDUCATION AT “NICHE” HEMATOPOIETIC STEM CELL DC BONE MARROW “NICHE” MHC HSC

114. Engineering A Bioartificial Heart NATURE MEDICINE Jan 13, 2008

115. Engineering A Bioartificial Heart NATURE MEDICINE Jan 13, 2008

116. STEM CELLS : FUTURE ODESSY...... 2004 2020 – 2040 2060 … CORD BLOOD STEM CELLS EMBRYONIC STEM CELLS ADULT STEM CELLS ? ? + + +

117. RONALD REAGAN CHRISTOPHER REEVE

118. 1 JULY 2005 WHAT DO WE KNOW & WHAT DON’T WE KNOW IN SCIENCE?

121. HEMATOPOIETIC STEM CELLS “ NICHE” “ Education Sites for Stem Cells”

122. KÖK HÜCRELERİ

123. BONE MARROW STEM CELLS : How Many ?? BM Windows XP

124. REPROGRAMMING OF SOMATIC CELLS TO THE EMBRYONIC STEM CELL SOMATIC CELLS ES-like cells SOMATIC CELLS ESC FUSION ES-like Cells

125. E.DONNALL THOMAS, M.D. Emeritus Professor of Medicine University of Washington Fred Hutchinson Cancer Research Center - Seattle, USA Nobel Laureate for his contributions to Hematopoietic Stem Cell Transplantation in hematologic disorders 1990 1990

127. MOUSE EMBRYONIC STEM CELLS Martin Evans’1981

128. 2007 Physiology and Medicine Oliver Smithies Univ North Carolina U.S.A. Mario Capecchi Univ of Utah U.S.A. Sir Martin Evans Cardiff Univ U.K. “ Contributions to Embryonic Stem Cell Biology and Development of Knock-Out Mouse Model to Understand Pathobiology of Diseases”

129. FUTURE OF STEM CELLS.....

130. DEVELOPMENT OF A BABY FROM EMBRYONIC STEM CELLS hESC

131. STAGES (Waves) OF HEMATOPOIESIS PRIMITIVE ERYTHROPOIESIS YOLK SAC Erytbroblasts, Macrophages Circulation / Fetal Liver TRANSIENT DEFINITIVE ERYTROPOIESIS Myelo-Erytrosid Progenitors (MEP) YOLK SAC LONG - TERM DEFINITIVE AGM YOLK SAC LARGE ARTERIES FETAL LIVER BONE MAROW Self-Renewing HSCs Lympho-Myelo-Erythroid FETAL LIVER CONCEPTION BIRTH

132. 0 20 DAY

133. BENCH  BED-SIDE

134. BREAKTHROUGHS of THE YEAR - 2007 SCIENCE December 21, 2007 318:1833 **Human Genetic Diversity and Personal “ Geneology” -Having One’s Own Genome ? *** How Adult Human Epithelial Cells could be re- programmed to act Like “Pluripotent Stem Cells” and like Embryonic Stem Cells... ** Progress in Climate Change Research and Impacts of global warming on Ecosystems and sea-ice.

135. STEM CELLS - MOUSE CLP T B NK CMP GMP G DC M MeP TR RBC HEMATOPOIETIC STEM CELLS (HSC) LONG-TERM STEM CELLS (LTSC) SHORT-TERM STEM CELLS (STSC) MULTIPOTENT PROGENITOR CELLS (MPC)

136. PROLIFERATION AND SELF-RENEWAL OF STEM CELLS IN “ NICHE” NICHE SIGNALLING TARGET CELLS WITHIN THE RANGE OF NICHE Ang-1/Tie-2 signalling and N-Cadherin contributes to NICHE - STEM CELL INTERACTION SC SC SC SIGNALLING CELLS EFFECTIVE RANGE OF SIGNALS

141. MEDICINE The Whole World Is Watching Hope—and anxiety—run high as the first clinical trial of embryonic-stem-cell therapy begins this summer. President Barack Obama’s executive order lifted restrictions on Embryonic Stem Cell Research NEWSWEEK March 30 , 2009

142. EMBRYONIC STEM CELLS ..Bench to Clinic ???

143. Endothelium vWF Alb/HNF1 Hepatocytes Neural cells Cardiac: No? Hemopoietic cells MESENCHYMAL STEM CELLS PDX1 TRITC Pancreas Endod *** *** *** CaPo4 Osteoblasts Chondroblasts Collagen II SM Myoblast Sm Myosin Sk Myoblast Sk Myosin

145. PROLIFERATION AND SELF-RENEWAL OF STEM CELLS IN “ NICHE” NICHE SIGNALLING TARGET CELLS WITHIN THE RANGE OF NICHE Ang-1/Tie-2 signalling and N-Cadherin contributes to NICHE - STEM CELL INTERACTION SC SC SC SIGNALLING CELLS EFFECTIVE RANGE OF SIGNALS

146. IN THE STEM CELL.... GENES mRNAs PROTEINS CELL FATE

147. MOUSE EMBRYO STEM CELL SC SC SC NEURONS Symmetric Division Asymmetric Division SC