Stem cell banking3

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stem cell by Dr / samir Mahmoud

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Stem cell banking3

  1. 1. WEB IRM Copyright 2002 Allyn & Bacon Customized by Marsha Blachman, LCSW
  2. 2. WEB IRM Copyright 2002 Allyn & Bacon Customized by Marsha Blachman, LCSW
  3. 3. Copyright 2002 Allyn & Bacon Dr /Samir Mahmoud Attia
  4. 4. Copyright 2002 Allyn & Bacon By Dr /Samir Mahmoud Attia
  5. 5. WEB IRM Copyright 2002 Allyn & Bacon Customized by Marsha Blachman, LCSW Aim of the work • This essay aim to clarify the technique of Banking of stem cells by different ways of collection and storing of blood stem cells
  6. 6. WEB IRM Copyright 2002 Allyn & Bacon Customized by Marsha Blachman, LCSW What Are Stem Cells ? Unspecialized, primitive Self- renewing cells that Can differentiate into cells with specific functions
  7. 7. WEB IRM Copyright 2002 Allyn & Bacon Customized by Marsha Blachman, LCSW
  8. 8. WEB IRM Copyright 2002 Allyn & Bacon Customized by Marsha Blachman, LCSW Classification of stem cell Embryonic, adult Hematopoietic, mesenchymal Tottipotent, Pleuripotentl Multipotent unipotent
  9. 9. WEB IRM Copyright 2002 Allyn & Bacon Customized by Marsha Blachman, LCSW Embryonic stem cells • Embryonic stem cells are only found naturally in the early stages of embryonic development and are totipotent i.e. they can form any type of adult cell or adult cell precursor.
  10. 10. WEB IRM Copyright 2002 Allyn & Bacon Customized by Marsha Blachman, LCSW Adult stem cells Have been found in: Brain Bone marrow Blood vessels Digestive tract Are multipotent, e.g., hematopoietic stem cells that form different blood components Skeletal muscle Skin Liver Umbilical cord
  11. 11. WEB IRM Copyright 2002 Allyn & Bacon Customized by Marsha Blachman, LCSW Hematopoietic stem cells A hematopoietic stem cell is a cell isolated from the blood or bone marrow that can renew itself. It can differentiate to many specialized cells.
  12. 12. WEB IRM Copyright 2002 Allyn & Bacon Customized by Marsha Blachman, LCSW Mesenchymal Stem Cells Mesenchymal cells are distinct from haematopoietic cells by being CD45 -. Mesenchymal cells are a mixed cells that are capable of supporting haematopoiesis and differentiating into endothelial, bone, muscle and neural cells.
  13. 13. WEB IRM Copyright 2002 Allyn & Bacon Customized by Marsha Blachman, LCSW
  14. 14. WEB IRM Copyright 2002 Allyn & Bacon Customized by Marsha Blachman, LCSW
  15. 15. WEB IRM Copyright 2002 Allyn & Bacon Customized by Marsha Blachman, LCSW 1. TOTIPOTENTIAL stem cells Derived from embryonic stem cells They can become any cell type They can renew themselves indefinitely Classification Of Stem Cells
  16. 16. WEB IRM Copyright 2002 Allyn & Bacon Customized by Marsha Blachman, LCSW 2. PLURIPOTENTIAL stem cells can grow into any cell type except totipotential stem cells. They cannot become an embryo. They can renew themselves indefinitely.
  17. 17. WEB IRM Copyright 2002 Allyn & Bacon Customized by Marsha Blachman, LCSW 3. MULTIPOTENTIAL stem cells Generate only closely related cells, e.g. blood cells such as white blood cells, red blood cells, lymphocytes, platelets… etc. They can renew themselves indefinitely.
  18. 18. WEB IRM Copyright 2002 Allyn & Bacon Customized by Marsha Blachman, LCSW 4. UNIPOTENTIAL (PROGENITOR) stem cells Produce only one cell type They also have the ability of self renewal, which distinguishes them from non-stem cells.
  19. 19. WEB IRM Copyright 2002 Allyn & Bacon Customized by Marsha Blachman, LCSW Sources of Stem Cells
  20. 20. WEB IRM Copyright 2002 Allyn & Bacon Customized by Marsha Blachman, LCSW Peripheral Blood Stem Cells (PBSC) The bloodstream is one source of stem cells, although not rich as a source as bone marrow. To have enough stem cells in a donor's bloodstream for a transplant, the donor is given a special drug called a "growth factor" (filgrastim is a drug that is commonly used for this purpose).
  21. 21. WEB IRM Copyright 2002 Allyn & Bacon Customized by Marsha Blachman, LCSW When enough stem cells are present in the bloodstream, the donor undergoes a process called apheresis. During an apheresis the blood stem cells are separated from the donor's blood, and the remaining blood goes back into the donor's bloodstream through a sterile needle.
  22. 22. WEB IRM Copyright 2002 Allyn & Bacon Customized by Marsha Blachman, LCSW Bone Marrow Stem Cells Bone marrow, a spongy tissue found inside larger bones, it is a rich source of blood stem cells. Approximately one liter of marrow is needed for a blood stem cell transplant, but the exact amount needed depends on the size of the patient. Marrow is removed during a surgical procedure. Sterile needles and syringes are used to remove
  23. 23. WEB IRM Copyright 2002 Allyn & Bacon Customized by Marsha Blachman, LCSW
  24. 24. WEB IRM Copyright 2002 Allyn & Bacon Customized by Marsha Blachman, LCSW Umbilical Cord Blood Stem Cells Umbilical cord blood is another rich source of stem cells. Months before the baby's birth, the mother signs an agreement to donate the umbilical cord blood when the baby is born. At birth, the cord blood unit is collected and taken to a cord blood bank, where it is tissue- typed, processed and stored frozen until needed for a transplant.
  25. 25. WEB IRM Copyright 2002 Allyn & Bacon Customized by Marsha Blachman, LCSW
  26. 26. WEB IRM Copyright 2002 Allyn & Bacon Customized by Marsha Blachman, LCSW Cord Blood Banking The first successful cord cell transplant to a sibling with Fanconi’s anemia took place in 1988. This proven utility of cord blood led to the establishment of cord blood banks. The first private and public banks were established in 1992.
  27. 27. WEB IRM Copyright 2002 Allyn & Bacon Customized by Marsha Blachman, LCSW • CB banking provides rapid availability of allogeneic donors for stem cell transplantation, little donor risk or attrition, low risk of transmitting infection, reduced or no risk of (AGVHD) .
  28. 28. WEB IRM Copyright 2002 Allyn & Bacon Customized by Marsha Blachman, LCSW Advantages and Disadvantages of Umbilical Cord Blood
  29. 29. WEB IRM Copyright 2002 Allyn & Bacon Customized by Marsha Blachman, LCSW Advantages of UCB 1) Cord blood has been found to possess more primitive cells. 2) Umbilical cord blood offers potential advantages,that infectious diseases is marked less than bone marrow or peripheral blood despite the possibility of congenital and perinatal transmission of infectious agents. for example, no cord bloods would be infected with Epstein Barr virus (EBV).
  30. 30. WEB IRM Copyright 2002 Allyn & Bacon Customized by Marsha Blachman, LCSW 3) CD34+ CD38- cells in umbilical cord blood also proliferate more rapidly in response to cytokine stimulation with IL-3, IL-6 and stem cell factor (SCF) and generate seven times more progenitor cells than do bone marrow .
  31. 31. WEB IRM Copyright 2002 Allyn & Bacon Customized by Marsha Blachman, LCSW 4) Umbilical cord blood has more haematopoietic stem cells per volume than peripheral blood or bone marrow. In addition, umbilical cord blood seems more tolerant of HLA mismatches; with less graft versus host disease 5) Cord blood is less risky to collect. The collection of cord blood following delivery is a harmless process that does not affect the mother or her newborn.
  32. 32. WEB IRM Copyright 2002 Allyn & Bacon Customized by Marsha Blachman, LCSW Disadvantages of UCB 1) A disadvantage of umbilical cord blood compared with donated adult bone marrow is that newborns may carry undiagnosed genetic diseases. 2) BM donors can be recalled for the same patient to provide a second donation of BM.
  33. 33. WEB IRM Copyright 2002 Allyn & Bacon Customized by Marsha Blachman, LCSW 3) BM donors can provide a medical history, which is relevant to the donation, at the time of donation. However, CB donors provide the medical history through their mothers. 4) that bone marrow contains more mesenchymal progenitor cells than CB.
  34. 34. WEB IRM Copyright 2002 Allyn & Bacon Customized by Marsha Blachman, LCSW
  35. 35. WEB IRM Copyright 2002 Allyn & Bacon Customized by Marsha Blachman, LCSW Bone Marrow/ Peripheral Blood Cord Blood -Donation requires surgery under general anesthesia (in B.M) -Donors feel discomfort and/or pain (in P.B). -Long-term consequences of growth factors used in peripheral blood stem cell donation posses no risk to mother or infant.
  36. 36. WEB IRM Copyright 2002 Allyn & Bacon Customized by Marsha Blachman, LCSW Large dose of stem cells. Rapid engraftment. Smaller dose of stem cells. Slower engraftment. After a formal search is begun, takes an average of 4 months to transplantion, if a donor is available. When a match is found, can take only a few days for confirmatory and special testing (may reach less than 24 hours in an emergency).
  37. 37. WEB IRM Copyright 2002 Allyn & Bacon Customized by Marsha Blachman, LCSW Potential donors may no longer be available or may have withdrawn consent. Donor must be found and retested to confirm the HLA typing and infectious disease results and to confirm that the donor is still willing and able to donate bone marrow . Once frozen, a cord blood unit is available until used.
  38. 38. WEB IRM Copyright 2002 Allyn & Bacon Customized by Marsha Blachman, LCSW Bone marrow must be used fresh (shelf-life measured in hours). Peripheral blood stem cells stored for short term (days to a few months). Frozen cord blood has been transplanted successfully up to 10 years in storage. Latent viral infection in the donor common (i.e. CMV > 50% in adult donors). Latent viral infection in the cord blood donor rare (i.e. CMV <1% ) Severe graft vs host disease (GvHD) common. GvHD less frequent and easier to treat.
  39. 39. WEB IRM Copyright 2002 Allyn & Bacon Customized by Marsha Blachman, LCSW Generally requires a perfect match between donor and recipient for HLA-A, -B . Additional factors HLA-C, -DQ and -DP)are needed to improve prognosis). HLA-mismatched cord blood transplantion are possible, making it easier to find a suitable match. No risk of transplanting a genetic disease. Cord blood has a small risk of transplantation rare unrecognized genetic disease.
  40. 40. WEB IRM Copyright 2002 Allyn & Bacon Customized by Marsha Blachman, LCSW Methods of umbilical cord blood collection • variety of potential collection methods (open, semi- closed or closed methods) have been proposed in order to optimize the collection volume and reduce the risks of microbial and maternal contamination.
  41. 41. WEB IRM Copyright 2002 Allyn & Bacon Customized by Marsha Blachman, LCSW
  42. 42. WEB IRM Copyright 2002 Allyn & Bacon Customized by Marsha Blachman, LCSW Donor selection • The medical history interview includes a review of the risk for HIV and hepatitis B and C, including skin piercing and blood transfusion, as well as the presence of infections that can be transmitted.
  43. 43. WEB IRM Copyright 2002 Allyn & Bacon Customized by Marsha Blachman, LCSW Collection of UCB • Method of Collections : • 1-In- utero: before delivery of placenta • 2-Ex- utero:After placental delivery
  44. 44. WEB IRM Copyright 2002 Allyn & Bacon Customized by Marsha Blachman, LCSW • Representation of the UCB collection system. The delivered placenta is placed onto the funnel-shaped supporting surface with the maternal side facing upward, the fetal side facing downward, and the umbilical cord passing through the central hollow cylinder of the supporting surface. The device lid is then closed, the pressure application system is turned on, and the UCB collection begins.
  45. 45. WEB IRM Copyright 2002 Allyn & Bacon Customized by Marsha Blachman, LCSW • Blood collection was made by puncturing the umbilical vein after the umbilical cord had been clamped, cut, and cleaned with an antiseptic solution, and drainage by gravity into a regular blood donation triple-bag system with the anticoagulant citrate phosphate dextrase (CPD) reduced to 25 mL
  46. 46. WEB IRM Copyright 2002 Allyn & Bacon Customized by Marsha Blachman, LCSW
  47. 47. WEB IRM Copyright 2002 Allyn & Bacon Customized by Marsha Blachman, LCSW
  48. 48. WEB IRM Copyright 2002 Allyn & Bacon Customized by Marsha Blachman, LCSW
  49. 49. WEB IRM Copyright 2002 Allyn & Bacon Customized by Marsha Blachman, LCSW Collection of UCB • Volume 80 – 120 mL. (30-40 mL..TNC > 6 x 10/9 cells) • Transport –By dry shipper( Refrigeration 4 C)
  50. 50. WEB IRM Copyright 2002 Allyn & Bacon Customized by Marsha Blachman, LCSW Processing of UCB • 24-36 hrs. post collection. – Max. 48 hrs. – Volume reduction • depletion of erythrocytes and/or plasma by: • HES (hydroxy ethyl starch) • Hespan. (1:5 ratio Hespan/UCB)
  51. 51. WEB IRM Copyright 2002 Allyn & Bacon Customized by Marsha Blachman, LCSW Processing of UCB • Analysis. – Cell Count/Diff, Viability (>70%), ABO, Rh, – CD34 +, Stem Cell Assay (post processing) – Sterility Testing (pre and post processing).
  52. 52. WEB IRM Copyright 2002 Allyn & Bacon Customized by Marsha Blachman, LCSW
  53. 53. WEB IRM Copyright 2002 Allyn & Bacon Customized by Marsha Blachman, LCSW Processing of UCB • Cryopreservation – 25 –50 mL Cryocyte Storage Container – 10% dimethyl sulfoxide (DMSO) – Control Rate Freeze Process • 1 C – 2 C /min
  54. 54. WEB IRM Copyright 2002 Allyn & Bacon Customized by Marsha Blachman, LCSW Most cord blood units were cryopreserved as whole blood donations with 10% DMSO and transfused immediately after thawing without any attempt to remove the cryoprotectant.
  55. 55. WEB IRM Copyright 2002 Allyn & Bacon Customized by Marsha Blachman, LCSW Thawing of cord blood units is usually performed by placing the cord blood bags in the gas phase of liquid nitrogen for 30 min followed by a 5-min exposure to room temperature. The bags are then thawed in a 37C water bath as rapidly as possible. Immediately after thawing, each cord blood unit is diluted with an equal volume of solution containing 5% human serum albumin and 10%
  56. 56. WEB IRM Copyright 2002 Allyn & Bacon Customized by Marsha Blachman, LCSW Dextran 40 in 0.9% NaCl, and sedimented for 10 min. The supernatant is then removed and sedimented cells are resuspended slowly in fresh albumin/ dextran solution. • This procedure removes the bulk of RBC ghosts, hemoglobin, and DMSO, thus reducing some of the risks associated with the transplantion procedure.
  57. 57. WEB IRM Copyright 2002 Allyn & Bacon Customized by Marsha Blachman, LCSW
  58. 58. WEB IRM Copyright 2002 Allyn & Bacon Customized by Marsha Blachman, LCSW
  59. 59. WEB IRM Copyright 2002 Allyn & Bacon Customized by Marsha Blachman, LCSW
  60. 60. WEB IRM Copyright 2002 Allyn & Bacon Customized by Marsha Blachman, LCSW
  61. 61. WEB IRM Copyright 2002 Allyn & Bacon Customized by Marsha Blachman, LCSW
  62. 62. WEB IRM Copyright 2002 Allyn & Bacon Customized by Marsha Blachman, LCSW Recommindations • 1-Written permission should be obtained during prenatal care and before the onset of labour. • 2-cord blood collection should not be done in complicated deliveries and the cord blood stem cell collection program should not alter routine practice for the timing of umbilical cord clamping.
  63. 63. WEB IRM Copyright 2002 Allyn & Bacon Customized by Marsha Blachman, LCSW • 3-condition such as leukemia or severe hemoglobinopathy may indicate the need for directed donor cord blood banking for sibling cord blood transplantation. • 4-preliminary data show encouraging results in cord blood stem cell transplantation for a variety of genetic, hematological and oncological diseases .
  64. 64. WEB IRM Copyright 2002 Allyn & Bacon Customized by Marsha Blachman, LCSW

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