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Dr R V S N Sarma MD MSc Consultant Physician & Chest Specialist

Stem Cell Research Advancements & Applications Special focus on Cord Blood Basics of Cloning and GE. Dr R V S N Sarma MD MSc Consultant Physician & Chest Specialist. www.drsarma.in. The Outline of Discussion. What are Stem Cells ? Why Stem Cells ? What are the sources of Stem Cells (SC) ?

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Dr R V S N Sarma MD MSc Consultant Physician & Chest Specialist

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  1. www.drsarma.in

  2. Stem Cell Research Advancements & ApplicationsSpecial focus on Cord BloodBasics of Cloning and GE Dr R V S N Sarma MD MSc Consultant Physician & Chest Specialist www.drsarma.in www.drsarma.in

  3. The Outline of Discussion • What are Stem Cells ? Why Stem Cells ? • What are the sources of Stem Cells (SC) ? • What are the methods of SC Production ? • How Cord Blood SCs are harvested ? • What are the applications of Stem Cells ? • Where are we now ? • Where do we go from here ? • Cord Blood Banking resources – www.drsarma.in

  4. Cell Therapy – Regenerative Medicine • Autologous (from the same individual) • Allogeneic (from a different individual) • Xenogeneic (from a different species) • Differentiated – specialized cells of same or other tissue type, such as heart, muscle, blood cells etc. • Undifferentiated or unspecialized or uncommitted cells such as stem cells. www.drsarma.in

  5. Different Types of Stem Cells • Totipotent (totally undifferentiated – potential to become any cell type including whole organism) • Example: The inner cell mass of embryo • Pluripotent (undifferentiated – potential to be any cell type but not an entire organism) • Emb Stem Cells (hESC), Emb Germ Cells (hEGC) • Multipotent (differentiated and yet undifferentiated) • The UCSC, BMSC, MSC – limited in potential Use of the stem cells raises ethical / legal concerns Wide variation in national policies for stem cell use www.drsarma.in

  6. Hierarchy of Stem Cells Multipotent www.drsarma.in

  7. What is special about Stem Cells ? • Capability of dividing and renewing themselves for long periods - almost immortal • Stem cells are unspecialized “uncommitted” cells. • Stem cells can give rise to specialized cells. Remain “uncommitted” until they receive signals from their environment to develop into specialized cells. • Stem Cells have Plasticity - Stem Cells from one tissue are able to give rise to cells of a completely different type of tissue www.drsarma.in

  8. Hectic Competition The ultimate in love www.drsarma.in

  9. 4 cell stage Late Day 2 Morula Day 3 2 cell stage Day 2 Blastocyst Day 5 Implantation Day 6 - 7 Fertilization Day 1 Week 1 of Gestation www.drsarma.in

  10. The Blastocyst – Day 5 The Blastocyst consists of Inner Cell Mass (embryoblast) Trophoblast Blastocele www.drsarma.in

  11. Week 2 of Gestation - Day 7.5 Uterine Artery Uterine Gland Syncitio-trophoblsat Blastocystic Cavity Uterine Epithelium Inner Cell Mass Cytotrophoblast Syncytiotrophoblast www.drsarma.in

  12. Week 2: Early Development Cytotrophoblast Amniotic Cavity Amnioblasts Inner Cell Mass : Epiblast Hypoblast Primary Yolk Sac Exocoelomic Membrane www.drsarma.in

  13. Embryonic Cell Types – at Gastrulation www.drsarma.in

  14. GENES www.drsarma.in

  15. Sexual Reproduction www.drsarma.in

  16. Cloning or Asexual Reproduction SCNT www.drsarma.in

  17. Parthenogenesis www.drsarma.in

  18. Embryonic Stem Cell Cultures www.drsarma.in

  19. Human Genetic Engineering www.drsarma.in

  20. Germline Engineering • Combines the application of • Stem Cell use • Genetic Engineering • Embryo Cloning and • To Produce Designer Babies www.drsarma.in

  21. In Vivo – Pluripotent Stem Cells www.drsarma.in

  22. Different Types of Stem Cells • Embryonic cells – by In Vitro Fertilization (IVF) • ESC and Embryonic Germ Cells (EGC) • Somatic cell nuclear transfer (SCNT) - Cloning • Used to clone ‘Dolly’ • Adult Stem Cells (ASC) – Best eg. BMT, MSC • Umbilical cord blood stem cells (UCSC) • Already differentiated to some degree ? • Can they be made to become pluripotent ? • Will they rapidly lose capacity to differentiate ? • Amniotic Fluid Stem Cells – AFSC – 9th Jan 2007 www.drsarma.in

  23. Embryonic Stem Cell (ESC) High malleability Potential for undesired development (teratomas) Infinite lifespan, unlimited supply High ethical burden Uncertain legal status Adult Stem Cells (ASC) Limited developmental potential Better behaved, easier to manage Lose their ability to proliferate or differentiate after a time in culture Less moral ambiguity Less legal controversy ESC versus ASC www.drsarma.in

  24. Methods of Stem Cell Generation IVF SCNT www.drsarma.in

  25. Stem Cell Maturation in Bone Marrow www.drsarma.in

  26. Alternatives to Embryonic Stem Cells from Living Human Embryos • From dead embryos • Less than 8 wks MTPs • Non-destructive biopsy • Bioengineered cells (genetically altered) • Reprogrammed adult somatic cells [but] Live human embryos hold most promise www.drsarma.in

  27. Stem Cell Differentiation Stem Cell Programming ‘RECIPE’ www.drsarma.in

  28. Therapeutic Applications of Stem Cells • Regenerative medicine – permanent repair of failing organs - e.g. • Cardiomyocytes for heart disease • Angiogenesis in CAD – ‘Auto bypass’ • Islet cells for diabetes • Neural cells for Parkinson’s • Blood cells for cancer • Chondrocytes for osteoporosis • Keratinocytes for burns www.drsarma.in

  29. Stem Cell Applications www.drsarma.in

  30. The Promise of Stem Cell Research www.drsarma.in

  31. Innovations of Cell Therapies www.drsarma.in

  32. Cell Therapies – Global Over View www.drsarma.in

  33. Estimated Market for CT versus IT www.drsarma.in

  34. Electron Micrographs of Stem Cells www.drsarma.in

  35. Programming of Stem Cells – ex vivo EMG of Stem Cell www.drsarma.in

  36. Stem Cells in CAD Cardiomyogenesis Coronary Angiogenesis www.drsarma.in

  37. Let us Understand The Enormous Power of the Umbilical Cord www.drsarma.in

  38. Umbilical Cord Blood Banking • Collection, Processing, Cryo-preservation of blood in the placenta and umbilical cord after the separation of baby • In-Utero collection, Ex-Utero collection • 45 disease are now treatable with Cord blood Stem Cells • These are not ‘experimental’ but ‘mainstream’ therapies • The stem cell concentration is 10 times higher than BM • For BMT overall only 25% get a suitable HLA match • Reduced GVH-Disease in the recipient • Painless and easy to collect, Immediately available • BMT costs 3-4 lakhs – Cord blood has lower cost www.drsarma.in

  39. UCSC Rx. for which diseases ? • Acute and Chronic Leukemias • Myelodystrophic syndromes • Stem Cell disorders – FA, PNHU, Aplastic Anemia • Myelo and Lympho proloferative Disorders • Liposomal storage Diseases, Plasma cell disorders • Phagocyte Disorders • Histiocytic Disorders, Platelet abnormalities • Other Malignancies – BC, Ewings, NB, RCC • RBC Abnormalities – Sickle cell, Beta Thalassemia • SCID – Congenital immune system disorders www.drsarma.in

  40. Promising Applications of UCSC • Parkinsonism • Alzheimer's Disease • Type 1 Diabetes Mellitus • Cardio-myopathies • Coronary revascularization for CAD • Retinoblastoma – Retinopathy • Burns and skin regeneration • Osteo and other degenerative arthritides • Cirrhosis, Hepatitis • Osteoporosis www.drsarma.in

  41. www.drsarma.in

  42. www.drsarma.in

  43. The Placental Barrier www.drsarma.in

  44. Why Use Cord Blood Stem Cells ? • Absolutely non-controversial • Absolutely simple to collect • More potent than Adult stem cells • Non-contaminated • Readily available • Both for vaginal and c-section deliveries • Low rates of rejection for transplants • Most importantly very high chances of HLA matching www.drsarma.in

  45. Stem Cell therapy centres in India • Tata Memorial Hospital, Mumbai • Adyar Cancer Centre, Madras • Apollo Specialty Hospital, Madras, • Apollo Hospital, Global Hospitals, NIMS, Hyderabad • Christian Medical College, Vellore • Narayana Hruduyalaya , Bangalore • R&R Army Hospital, New Delhi • AIIMS , New Delhi • Inlaks Hospital, Pune • Armed Forces Medical College, Pune • Sanjay Gandhi PGIMS, Lucknow www.drsarma.in

  46. Storage of Umbilical Bloodfor Stem Cells • Commercial enterprise (Cordbank) stores umbilical stem cells from birth for future autologous use. “Saving your baby’s umbilical cord stem cells could save your baby’s life” • Others advocate for a public cord blood bank www.drsarma.in

  47. Cord Blood Collection www.drsarma.in

  48. Thawing Method • Based upon COBLT (Cord Blood Transplantation Study) Method • Used for thawing cord blood products received from COBLT Banks frozen in MedSep (80/20) bags www.drsarma.in

  49. Cord Blood Preservation Under liquid N2 at – 192o C www.drsarma.in

  50. Preparation 2 Hours prior to thaw: • Review product paperwork • Gather supplies and reagents • Prepare wash media (Dextran 40 and human serum albumin). • Prepare wash bag setup: MedSep Transplant Set+ 300 mL plastic transfer bag. www.drsarma.in

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