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II. Blood and Blood Components

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  1. II. Blood and Blood Components Terry Kotrla, MS, MT(ASCP)BB Spring 2010

  2. Goals Of Blood Collection • Maintain viability and function • Prevent physical changes • Minimize bacterial contamination

  3. Anticoagulants Preservative Solutions • Anticoagulants prevent blood clotting • Preservatives provide nutrients for cells • Heparin • Rarely if ever used anymore • Anticoagulant ONLY • Transfuse within 48 hours, preferably 8

  4. Anticoagulants

  5. Additive Solution • Primary bag with satellite bags attached. • One bag has additive solution (AS) • Unit drawn into CPD anticoagulant

  6. Additive Solution • Remove platelet rich plasma within 72 hours • Add additive solution to RBCs, ADSOL, which consists of: • Saline • Adenine • Glucose • Mannitol • Extends storage to 42 days • Final hematocrit approximately 66%

  7. Changes Occur During Storage • Shelf life = expiration date • At end of expiration must have 75% recovery • At least 75% of transfused cells remain in circulation 24 hours AFTER transfusion

  8. Storage Lesion • Biochemical changes which occur at 1-6C • Affects oxygen dissociation curve, increased affinity of hemoglobin for oxygen. • Low 2,3-DPG, increased O2 affinity, less O2 released. • pH drops causes 2,3-DPG levels to fall • Once transfused RBCs regenerate ATP and 2,3-DPG • Few functional platelets present • Viable (living) RBCs decrease

  9. Plasma hemoglobin Plasma K+ Na+ K+ Viable cells pH ATP 2,3-DPG Plasma Na+ Helps release oxygen from hemoglobin (once transfused, ATP & 2,3-DPG return to normal)

  10. Storage Lesion • Significant for infants and massive transfusion. • Other biochemical changes • ATP decreases • Potassium increases • Sodium decreases • Plasma hemoglobin increases

  11. Preparation of Components • Collect unit within 15 minutes to prevent activation of coagulation system • Draw into closed system – primary bag with satellite bags with hermetic seal between. • If hermetic seal broken transfuse within 24 hours if stored at 1-4C, 4 hours if stored at 20-24C

  12. Preparation of Components • Centrifuge – light spin, platelets suspended • Remove platelet rich plasma (PRP) • Centrifuge PRP heavy spin • Remove platelet poor plasma • Freeze plasma solid within 8 hours • Thaw plasma at 1-4C – precipitate forms • Centrifuge, express plasma leaving cryoprecipitate. Store both at -18C • RBCs – CPD – 21 days, ADSOL – 42 days – 1-6C

  13. Preparation of Components • Summary – One unit of whole blood can produce: • Packed RBCs • Fresh frozen plasma (FFP) • Cryoprecipitate (CRYO) • Single donor plasma (SDP) – cyro removed • Platelets – terms PC (platelet concentrate) OR RD PC (random donor platelet concentrate)

  14. Preparation of Components • Sterile docking device joins tubing • Used to add satellite bags to maintain original expiration of component • May be used to pool components

  15. Blood Component General Information • Blood separated into components to specifically treat patients with product needed • Advantages of component separation • Allow optimum survival of each component • Transfuse only component needed

  16. Blood Component General Information • Transfusion practice • Transfusion requires doctor’s prescription • All components MUST be administered through a filter • Infuse quickly, within 4 hours • D (Rh) neg require D neg cellular products • ABO identical preferred, ABO compatible OK • “Universal donor” – RBCs group O, plasma AB

  17. Blood Component General Information • Fresh Whole Blood • Blood not usually available until 12-24 hours • Candidates • Newborns needing exchange transfusion • Patients requiring leukoreduced products

  18. Blood Component General Information • Summary of storage temperatures: • Liquid RBCs 1-6C • Platelets, Cryo (thawed) and granulocytes 20-24C (room temperature) • ANY frozen plasma product ≤ -18C • ANY liquid plasma product EXCEPT Cryo 1-6C

  19. Blood Components • Cellular • Red blood cell products • Platelets • Granulocytes • Plasma • FFP • Cryoprecipitate

  20. Products With Red Cells

  21. Whole Blood • Clinical indications for use of WB are extremely limited. • Used for massive transfusion to correct acute hypovolemia such as in trauma and shock, exchange transfusion. • RARELY used today, platelets non-functional, labile coagulation factors gone. • Must be ABO identical.

  22. Changes in Stored Blood

  23. Red Blood Cells, Packed (PRBC) • Used to treat symptomatic anemia and routine blood loss during surgery • Hematocrit is approximately 80% for non-additive (CPD), 60% for additive (ADSOL). • Allow WB to sediment or centrifuge WB, remove supernatant plasma.

  24. Leukocyte Reduced Red Cells (LR-RBC) • Leukocytes can induce adverse affects during transfusion, primarily febrile, non-hemolytic reactions. • Reactions to cytokines produced by leukocytes in transfused units. • Other explanations to reactions include: immunization of recipient to transfused HLA or granulocyte antigens, micro aggregates and fragmentation of granulocytes. • Historically, indicated only for patients who had 2 or more febrile transfusion reactions, now a commonly ordered, popular component. • “CMV” safe blood, since CMV lives in WBCs. • Most blood centers now leukoreduce blood immediately after collection. • Bed side filters are available to leukoreduce products during transfusion.

  25. Leukocyte Reduction

  26. Washed Red Blood Cells (W-RBCs) • Washing removes plasma proteins, platelets, WBCs and micro aggregates which may cause febrile or urticarial reactions. • Patient requiring this product is the IgA deficient patient with anti-IgA antibodies. • Prepared by using a machine which washes the cells 3 times with saline to remove and WBCs. • Two types of labels: • Washed RBCs - do not need to QC for WBCs. • Leukocyte Poor WRBCs, QC must be done to guarantee removal of 85% of WBCs. No longer considered effective method for leukoreduction. • e. Expires 24 hours after unit is entered.

  27. Cell Washer to Prepare Washed Cells

  28. Frozen Blood

  29. Red Blood Cells Frozen; Red Blood Cells Deglycerolized (D-RBC) • Blood is frozen to preserve: rare types, for autologous transfusion, stock piling blood for military mobilization and/or civilian natural disasters. • Blood is drawn into an anticoagulant preservative. • Plasma is removed and glycerol is added. • After equilibration unit is centrifuged to remove excess glycerol and frozen. • Expiration • If frozen, 10 years. • After deglycerolization, 24 hours. • Storage temperature • high glycerol -65 C. • low glycerol -120 C, liquid nitrogen.

  30. Red Blood Cells Frozen; Red Blood Cells Deglycerolized (D-RBC) • Thaw unit at 37C, thawed RBCs will have high concentration of glycerol. • A solution of glycerol of lesser concentration of the original glycerol is added. • This causes glycerol to come out of the red blood cells slowly to prevent hemolysis of the RBCs. • After a period of equilibration the unit is spun, the solution is removed and a solution with a lower glycerol concentration is added. • This procedure is repeated until all glycerol is removed, more steps are required for the high glycerol stored units. • The unit is then washed.

  31. Rejuvenated Red Blood Cells • A special solution is added to expired RBCs up to 3 days after expiration to restore 2,3-DPG and ATP levels to prestorage values. • Rejuvenated RBCs regain normal characteristics of oxygen transport and delivery and improved post transfusion survival. • Expiration is 24 hours or, if frozen, 10 years

  32. Platelet Products

  33. Platelets (PLTS), Platelet Concentrate (PC) or Random Donor Platelet Concentrate (RD-PC) • Used to prevent spontaneous bleeding or stop established bleeding in thrombocytopenic patients. • Prepared from a single unit of whole blood. • Due to storage at RT it is the most likely component to be contaminated with bacteria. • Therapeutic dose for adults is 6 to 10 units. • Some patients become "refractory" to platelet therapy. • Expiration is 5 days as a single unit, 4 hours if pooled. • Store at 20-24 C (RT) with constant agitation. • D negative patients should be transfused with D negative platelets due to the presence of a small number of RBCs.

  34. Preparation of platelet concentrate Plasma RBCs PRP Platelet concentrate

  35. Platelets (PLTS), Platelet Concentrate (PC) or Random Donor Platelet Concentrate (RD-PC) • One bag from ONE donor • Need 6-10 for therapeutic dose

  36. Pooling Platelets • 6-10 units transferred into one bag • Expiration = 4 hours

  37. Platelets Pheresis, Apheresis Platelet Concentrate, Single Donor Platelet Concentrate (SD-PC) • Used to decrease donor exposure, obtain HLA matched platelets for patients who are refractory to RD-PC or prevent platelet refractoriness from occurring. • Prepared by hemapheresis, stored in two connected bags to maintain viability. • One pheresed unit is equivalent to 6-8 RD-PC. • Store at 20-24 C (RT) with agitation for 5 days, after combining, 24 hours • D negative patients should be transfused with D negative platelets due to the presence of a small number of RBCs

  38. Apheresis

  39. Apheresis

  40. Platelets Pheresis • One bag (unit) from one donor • One unit is a therapeutic dose • Volume approximately 250 ccs

  41. Granulocytes Lymphocyte Monocyte Neutrophils Eosinophils Basophils

  42. Granulocytes • Primary use is for patients with neutropenia who have gram negative infections documented by culture, but are unresponsive to antibiotics. • Therapeutic efficacy and indications for granulocyte transfusions are not well defined. • Better antimicrobial agents and use of granulocyte and macrophage colony stimulating factors best for adults, best success with this component has been with babies • Daily transfusions are necessary. • Prepared by hemapheresis. • Expiration time is 24 hours but best to infuse ASAP. • Store at 20-24 C.

  43. Plasma Components

  44. Fresh Frozen Plasma – Volume 200-250cc

  45. Fresh Frozen Plasma (FFP) • Used to replace labile and non-labile coagulation factors in massively bleeding patients OR treat bleeding associated with clotting factor deficiencies when factor concentrate is not available. • Must be frozen within 8 hours of collection. • Expiration • frozen - 1 year stored at <-18 C. • frozen - 7 years stored at <-65 C.thawed - 24 hours

  46. Fresh Frozen Plasma (FFP) • Storage temperature • frozen -18 C, preferably -30 C or lower • thawed - 1-6 C • Thawed in 30-37C water bath or FDA approved microwave • Must have mechanism to detect units which have thawed and refrozen due to improper storage. • Must be ABO compatible

  47. Plasma, Liquid Plasma, Recovered Plasma and Source Plasma • Used to treat patients with stable clotting factor deficiencies for which no concentrate is available or for patients undergoing therapeutic plasmapheresis. • Prepared by separating the plasma from the RBCs on or before the 5th day after expiration of the whole blood. • Once separated can: • Freeze, store at -18 C for 5 years • If not frozen, called liquid plasma, store at 1-6 C for up to 5 days after expiration of WB. • Once FFP is one year old can redesignate as Plasma, expiration is 5 years.

  48. Pooled Plasma/Solvent Detergent Treated • Most recently licensed product. • Prepared from pools of no more than 2500 units of ABO specific plasma frozen to preserve labile coagulation factors. • Treated with chemicals to inactivate lipid-enveloped viruses. • Contains labile and non-labile coagulation factors but lacks largest Von Willebrand’s factor multimers. • Used same as FFP.Safety concerns • Decreases disease transmission for diseases tested for. • Doesn’t inactivate viruses with non-lipid envelopes: parvo virus B19, hepatitis A, and unrecognized pathogens

  49. Cryoprecipitate (CRYO), Factor VIII or Anti-Hemophilic Factor (AHF) • Cold insoluble portion of plasma that precipitates when FFP is thawed at 1-6C. • Cryoprecipitate contains high levels of Factor VIII and Fibrinogen, used for treatment of hemophiliacs and Von Willebrands when concentrates are not available. • Used most commonly for patients with DIC or low fibrinogen levels. • A therapeutic dose for an adult is 6 to 10 units. • Can be prepared from WB which is then designated as "Whole Blood Cryoprecipitate Removed" or from FFP • Plasma is frozen. • Plasma is then thawed at 1-6 C, a precipitate forms. • Plasma is centrifuged, cryoprecipitate will go to bottom. • Remove plasma, freeze within 1 hour of preparation