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Stem Cells

Stem Cells. Embryonic stem cells Adult (Somatic) stem cells. ES cells line. Pluripotentiality property Immortal property. Nucleus transfer. Fusion of somatic cells and embryonic cells. Embryonic stem cell (ES) Endoderm Mesoderm Ectoderm Germ cell line

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Stem Cells

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  1. Stem Cells Embryonic stem cells Adult (Somatic) stem cells

  2. ES cells line Pluripotentiality property Immortal property

  3. Nucleus transfer Fusion of somatic cells and embryonic cells

  4. Embryonic stem cell (ES) Endoderm Mesoderm Ectoderm Germ cell line Mesenchymal stem cell (MSC) HSC adipocyte Gonad Death muscle tendon stromal osteoctye neuronal

  5. Adult (Somatic) Stem Cells Hematopoietic stem cells (HSC) Mesenchymal stem cells (MSC) Neural stem cells Hepatic stem cells etc.

  6. HSC Transplantation What is stem cell? CD 34+ ? (HSC) CD 34- Could it be stem cell? Is phenotype alone enough to identify stem cell?

  7. HSC Transplantation Autologous Allogeneic Syngeneic Xenogeneic

  8. Human Leukocyte Antigen (HLA) HLA class I A B C HLA class II DR DP DQ HLA match = 6 antigens (A B DR)

  9. HSC Transplantation Allogeneic HLA-matched related HLA-mismatched related: Haploidentical (3 Ags mismatched) HLA-matched unrelated HLA-mismatched unrelated

  10. HSC Transplantation Sources of Stem Cell Bone Marrow Peripheral Blood Stem Cell Cord Blood Fetal liver

  11. Differences among various sources of stem cells BM PBSC CB FL Engraftment ++ +++ + ? GVHD + ++ +/- -/? HLA matching yes yes yesno

  12. Process of HSC transplantation Conditioning Stem cell infusion GVHD prophylaxis Engraftment Immune tolerance

  13. Donor Recipient T cell NK cell T cell NK cell Stem cell Stem cell

  14. Indications for HSC transplant Malignant diseases: leukemia/lymphoma and hematologic malignancies and solid tumors Non malignant diseases: Bone marrow failure: aplastic anemia Thalassemia Primary immune deficiency Neurometabolic disease Metabolic bone disease Other genetic diseases Autoimmune disease

  15. Pediatric HSC Transplant at Ramathibodi: 1989-2005 Total patient 150 patients Autologous 30 patients Allogeneic 120 patients

  16. Pediatric HSC Transplant at Ramathibodi: 1989-2005 Age ranged 1 – 22 yr (median; 10 yr)

  17. Thalassemia α thalassemia disease Hb bart ( _ _ / _ _ ) Hb H disease ( _ _ /_ α ) β thalassemia disease Homozygous β thalassemia β thalassmia / Hb E

  18. Treatments in Thalassemia Palliative treatment Blood transfusion Iron chelation Splenectomy Curative treatment Stem cell transplant

  19. Pediatric Stem Cell Transplant in Thalassemia at Ramathibodi: 1989-2005 Thalassemia (n= 48 pts) Donor of stem cell Matched and mismatched related 25 pts Matched and mismatched unrelated 21 pts Haploidentical 2 pts Source of stem cell BMT and PBSCT 44 pts Cord blood 4 pts

  20. Figure 1 : Thalassaemia free survival for all patients N=49 - 1.0 0.9 0.8 0.7 0.6 0.5 0.4 0.3 0.2 0.1 0.0 78% Survival probability

  21. Figure 2 : Overall survival for all patients n=49 89% - 1.0 0.9 0.8 0.7 0.6 0.5 0.4 0.3 0.2 0.1 0.0 Survival probability

  22. Figure 3 : Thalassaemia free survival in related and unrelated transplantation groups - 1.0 0.9 0.8 0.7 0.6 0.5 0.4 0.3 0.2 0.1 0.0 82% n=28 p=0.42 70% n=21 Survival probability Related donor group Unrelated donor group

  23. Figure 4 :Overall survival in related and unrelated donor transplantation groups 92% n=28 - 1.0 0.9 0.8 0.7 0.6 0.5 0.4 0.3 0.2 0.1 0.0 p=0.43 82% n=21 Survival probability Related donor group Unrelated donor group

  24. Mesengenic Process :Caplan AI.Clin Plas Surg 1994;21(3):429-435.

  25. Adult stem cells and plasticity

  26. Day 2 Day 10

  27. Osteogenic Differentiation • MSCs were cultured for 16 days in the presence of • Dexamethazone • L-ascorbic acid-2-phosphate • Ascorbic acid • Beta-glycerophosphate • Detection of APase activity, formation of mineralized matrix :Jaiswal N et al. J Cell Biochem 1997;64:295-312.

  28. Induction of Osteoblast differentiation from Human Mesenchymal Stem Cells by Dexamethasone สินี ดิษฐบรรจง หน่วยโรคไต คณะแพทยศาสตร์ รพ.รามาธิบดี

  29. Methods hMSC passage 11-15 at confluency Dexamethasone 10-7 M L-ascorbate-2-phosphate 10 μM -glycerophosphate 10 mM

  30. Day 0 5 10 15 0 5 10 15 0 5 10 15 GAPDH Cbfa-1 Coll-1 Cbfa-1 Expression by RT-PCR 0 5 10 15 0 5 10 15 0 5 10 15

  31. Alkaline Phosphatase Enzyme Activity

  32. Day 7 14 21 28 Von Kossa Staining

  33. Stem cell transplant in Duchenne muscular dystrophy Hematopoietic stem cell and mesenchymal stem cell transplant Evaluation FISH XX/XY Dystrophin protien Dystrophin mutation analysis

  34. Dystrophin Staining Pre-transplantation

  35. Dystrophin Staining 1 year post transplantation

  36. Genetic control of chondrocyte and osteoblast differentiation

  37. 18 srRNA MSC (normal) CD 34+CB CD105 (thal)

  38. Study of Mechanism Osteoporosis Renal tubular acidosis Thalassemia

  39. Ex vivo umbilical cord blood expansion by co-culturing with IL-1 treated mesenchymal stem cells Day 7 Day 14 Day 21 MNC (fold) 6.0 22.4 33.9 CD34+ (fold) 1.1 52.0 89.0

  40. MSC: Potential Precursors for Tumor Stroma Target-Delivery Vehicles for Anticancer Agents Interferon IL-2 etc.

  41. Tolerance Induction in Organ Transplantation by Stem Cell Transplantation

  42. Donor Recipient T cell NK cell T cell NK cell Stem cell Stem cell

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