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Blood component preparation
Blood can be separated into components to meet most transfusion needs while minimizing risks. Effective separation requires centrifugation based on differences in specific gravity of components. Common components prepared include packed red blood cells, platelet concentrates, fresh frozen plasma, and granulocyte concentrates. Preparation of blood components allows for optimal use of donated blood by providing only the required constituent to the patient in need.
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Blood component preparation
- 1. BLOOD COMPONENTS INDICATIONS,PREPARATION OF BLOODCOMPONENTS SUNIL KUMAR.P Department of Haematology St.John’s Medical College Hospital Bangalore 1SUNIL KUMAR.P
- 2. INTRODUCTION • These dayseffective blood transfusion depends upon the availability of different blood components. • These components are used in separately or in combinations, can meet most patients transfusions need and keeping the risk of transfusion to a minimum. 2SUNIL KUMAR.P
- 3. WHY THERE ISNEED FOR COMPONENT SEPARATION • 1. Separation of blood in to components allows optimal survival of each constituents. • 2. component preparation allows transfusing only specific blood component that the patient requires. • 3. Transfusion of only specific constituent of blood needed avoids the use of unnecessary component, which could be contraindicated in a patient 3SUNIL KUMAR.P
- 4. • 4.By usingblood components, several patients can be treated with the blood from one donor, giving the optimal use of every unit of donated blood. • 5. use of blood components, supplements blood supply- adds to the blood inventory 4SUNIL KUMAR.P
- 5. BLOOD COMPONENTS Cellular components: • Red cell concentrate • Leucocytes-Reduced Red cells • Platelet Concentrate • Leukocytes-Reduced Platelet Concentrate • Platelet Apheresis • Granulocytes Apheresis 5SUNIL KUMAR.P
- 6. Plasma components • Freshfrozen plasma • Single donor plasma • Cryoprecipitate • Cryo-poor plasma • Plasma derivatives Albumin 5% & 25% 6SUNIL KUMAR.P
- 7. Plasma Protein Fractions •Factor VIII concentrate Immunoglobulins • Fibrinogen Other coagulation factors 7SUNIL KUMAR.P
- 8. PREPARATION OF BLOOD COMPONENTS •Equipment: • Freezer - 40°C and - 70°C • Blood bank refrigerator 2-6°C • Refrigerated centrifuge with swing out head and oval cups • Laminar flow • Weighing scale • Dielectric sealer or aluminium clips & sealer • Stripper • Water bath 37°C or Plasma Defroster (MicroWave) 8SUNIL KUMAR.P
- 9. • Platelet reciprocator(platelet incubator with agitator) • Cryoprecipitate bath 4°C 9SUNIL KUMAR.P
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- 15. Preparation of bloodcomponents is possible due to : • Multiple Plastic packs system • Refrigerated centrifuge • Different specific gravity of cellular components • Red cells specific gravity 1.08 - 1.09 • Platelet specific gravity 1.03 - 1.04 • Plasma specific gravity 1.02 - 1.03 15SUNIL KUMAR.P
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- 17. Methods of preparation •There are various methods to separate these components and the yield and quality of component depends upon the methods applied. Various methods used are: • a)Gravity separation • b)Low and high speed centrifugation • c)Apheresis by cell separator 17SUNIL KUMAR.P
- 18. WHOLE BLOOD • 350-450ml of blood • 49-63ml of anticoagulant solution • Hct- 36-44% • No components removed • Stored at 1- 6 degree celsius • Shelf life : • Citrate –phosphate –Dextrose(CPD)- 21days • CPDA (adenine) – 35 days • AS-1, AS-3, AS-5 – 42 days 18SUNIL KUMAR.P
- 19. • Increase Hbby 1 g/dl • Rate of Blood Transfusion -3ml/kg/hr 19SUNIL KUMAR.P
- 20. WHOLE BLOOD Indications •Acute active blood loss with hypovolemia • Exchange transfusion Contradictions Risk of volume overload • chronic anemia • Incipent cardiac failure 20SUNIL KUMAR.P
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- 22. WHOLE BLOOD Drawbacks •After storage for 24 hours platelets and wbc are non functional • Factor V and Viii decrease with storage • Fluid overload 22SUNIL KUMAR.P
- 23. Preparation of PackedRBCs Principle • RBCs are obtained by removal of supernatant plasma from centrifuged whole blood Preparation • 1. Collect appropriate volume of donor blood in CPDA double or triple bag. • 2. Store at 2-6C till processed. • 3. Place bags in the buckets of refrigerated centrifuge and balance the opposite bags accurately. • 4. Centrifuge at heavy spin (5000 x g) for 5 minutes at 2-6C. 23SUNIL KUMAR.P
- 24. • 5. Expressapproximately 3/4 of the plasma into the satellite bag. • 6. Double seal the tube between primary and satellite bags with plasma Separate the satellite bag with plasma and keep at -30°C or below. • 7. Keep the red cells at 2-6°C. 24SUNIL KUMAR.P
- 25. Indications • Patient needurgent operation and has Hb < 10g/dl • Anticipated surgical blood loss > 1000 ml • Other acute Loss of blood • Replacement Fluid blood loss < 20% of blood vol. • Anemia associated with incipient/established cardiac failure • Hb value < 6 g/dl • Patients approaching delivery and has Hb value < 7g/dl • In hereditary hemolytic anemias and betathalassaemia major. 25SUNIL KUMAR.P
- 26. Packed RBCs • Shelflife – 35 days • Storage temp- 2 – 6 Degree celsius • QC Requirements: PCV 80% (Range 65-80%) • Volume- 250- 300 ml Contents : • red cells- 65-80%, • Plasma 20- 35%, • Some platelets, • white blood cell and anticoagulant preservative solutions Effect – 1 unit RBC raise Hct by 3% and hb by 1g/dl 26SUNIL KUMAR.P
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- 28. Guidelines for Paediatricred cell transfusions Anaemia Infants within first 4 months of life • Hb< 13.0 g/dL and severe pulmonary disease • Hb < 10.0 g/dL and moderate pulmonary disease • Hb< 13.0 g/dL and severe cardiac disease • Hb<10.0 g/dL and major surgery • Hb < 8.0 g/dL and symptomatic anemia • Acute blood loss >10% of total blood volume. 28SUNIL KUMAR.P
- 29. Guidelines for Paediatricred cell transfusions Infants >4 months & children • Acute loss > 25% circulating blood volume • Hb < 8.0 g/dL in perioperative period • Hb < 13.0 g/dL and severe cardiopulmonary disease • Hb < 8.0 g/dL and marrow failure • Hb < 8.0 g/dL and symptomatic chronic anemia 29SUNIL KUMAR.P
- 30. IRRADIATED RED BLOODCELLS • Gamma radiated to kill the lymphocytes • The lack of T cells prevents graft versus host reaction Indications: • Severely imunocompromised patients • Lymphoma patients • Stem cell/marrow transplants • Intrauterine transfusion • Neonates undergoing exchange transfusion • Hodgkin Disease 30SUNIL KUMAR.P
- 31. LEUCOCYTE REDUCED RBCS Methodsof the preparation of Leucocyte- Reduced Red cells • Centrifugation and removing of buffy coat • Filtration • Washing of red cells with saline • Freezing and thawing of red cells 31SUNIL KUMAR.P
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- 33. LEUCOCYTE REDUCED RBCS INDICATIONS •Multi transfused patients like thalasemia • Leukemia • Aplastic anemia • Immunosupressed • Immunodeficient • Prevention of recurrent FNHTRs • Prevention or delay of primary alloimmunization to HLA antigen • Prevention of CMV transmission in at risk individual 33SUNIL KUMAR.P
- 34. FROZEN/DEGLYCEROLIZED RED CELLS Indications •The provision of rare blood lacking common antigens for patients with corresponding antibodies. • Autologous units of patients with multiple red cells alloantibodies stored for future surgery. • IgA deficient patient with antibodies to IgA • It can be used to prevent Febrile non hemolytic transfusion reactions( FNHTRs ) 34SUNIL KUMAR.P
- 35. GRANULOCYTE CONCENTRATE • Obtainedby apheresis from family members for administration in cancer patients • Contains 109granulocyte • Pretreatment with recombinant G-CSF and dexamethasone can yield 4-8x 108granulocyte • Stored at 24 degree celsius • Infuse within 24 hours of collection 35SUNIL KUMAR.P
- 36. GRANULOCYTE CONCENTRATE • Granulocyteconcentrate can be prepared by • 1. Single donor unit • 2. Leukapheresis by blood cells separator 36SUNIL KUMAR.P
- 37. INDICATIONS The role ofgranulocytes transfusion has decreased with • The availability of new and better antibiotics • The advent of recombinant granulopoeitic growth factors e.g. granulocyte stimulating factor . 37SUNIL KUMAR.P
- 38. Adverse effects ofgranulocytes transfusion •However the granulocytes transfusion are still used at times with success in the following conditions: • Septicemia not responding to antibiotics.. • Neutrophil count less than 0.50 x 109/L (500 / mm3), having gram-negative infection which fail to respond with antibiotics 38SUNIL KUMAR.P
- 39. • Temporary bonemarrow depression for 1 - 2 weeks • Infections in patients undergoing chemo / radio therapy for neoplastic diseases 39SUNIL KUMAR.P
- 40. PLATELET RICH PLASMA(PRP) Procedure: • 1. Collect 450 ml blood by a clean, single venipunctuer into 450 ml CPDA or Adsol / SAGM triple bags system. • 2. Keep the blood bag at room temperature (20-22°C) before preparing platelets, for not more than 6 hours. Do not chill at any time before or during platelet separation. • 3. Centrifuge the blood bags at 20-24°C at light spin for appropriate time (2000 x g for 3 minutes). • 4. Separate 4/5 of the platelet rich plasma (PRP) into one satellite bag 40SUNIL KUMAR.P
- 41. PLATELET CONCENTRATE • Shelflife: 1 day if no storage cabinet 5 days in platelet incubator and agitator • Storage temp :20-24 degree celsius • Q.C Requirements: to be prepared within 6hr of collection and ph should be >6.2 • volume: 30 to 50 ml • Contents: 5.5x1010 unit/bag 41SUNIL KUMAR.P
- 42. Indications Platelet countis < 5000 / µl regardless of clinical condition Effect: increases platelet count by 10000/ µl per unit 42SUNIL KUMAR.P
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- 44. PROCEDURE 1. Centrifuge thebag with PRP and another satellite bag at 20-40°C at ‘heavy spin’ for appropriate time e.g. 5000xg for 5 minutes. 2. Express supernatant platelet - poor plasma into another empty satellite bag. 44SUNIL KUMAR.P
- 45. SINGLE DONOR PLATELETS •Obtained by plasmapheresis technique • 6 to 8 times more platelets as in random donor unit • Larger volumes and HLA compatibility results in increase of 30 to 60 k • Leucoreduced because of apheresis technique • ABO Matched platelets preferred 45SUNIL KUMAR.P
- 46. FRESH FROZEN PLASMA •FFP is plasma obtained from a) a single donor either by normal donation or b) by plasmapheresis It contains all coagulation factors and a) plasma proteins Factor viii – 0.6 IU/ml b) Factor ix – 0.9 IU/ml c) Fibrinogen- 250-300 mg/bag Proteins – albumin, globulin etc 1 IU/kg of factor viii or factor ix raises factor viii in plasma by 2% and factor ix by 1% d) Volume: 200 – 250ml e) Shelf life- 1 year Storage temp(-20 to -70) degree or below 46SUNIL KUMAR.P
- 47. Procedure • 1. Collectappropriate volume of blood in 350- 450 ml CPDA double bags systems or 450 ml SAGM/Adsol triple bags system. • 2. Centrifuge at heavy spin (5000 x g for 5 minutes) at 4°C . • 3. Express approximately four fifth of the plasma into a satellite bag. 47SUNIL KUMAR.P
- 48. • Double sealthe tube between primary bag and the satellite bag having plasma with metal clips or dielectrical sealer. • Separate the satellite bag having plasma 48SUNIL KUMAR.P
- 49. Indications of FreshFrozen Plasma • Actively bleeding and multiple coagulation factors deficiencies • Liver diseases • Disseminated intravascular coagulation (D1C) • Coagulopathy in massive transfusion • TTP 49SUNIL KUMAR.P
- 50. • Antithrombin IIIdeficiency • Congenital or acquired coagulation factor deficiency • Use of FFP in conjunction with red cells has largely replaced the transfusion of fresh blood. • Deficiency of Factors II, VII, IX,, and X 50SUNIL KUMAR.P
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- 52. SINGLE DONOR PLASMA •Prepared from stored blood • Poor in coagulation factors • Cannot be used in correction of coagulation factor deficiencies • Effective as a volume expander 52SUNIL KUMAR.P
- 53. CRYOPRECIPITATE • Shelf life:frozen – 1 year • Thawed : 6 hours • Storage temp : frozen -20 degree or less • thawed : 37 degree • Volume- 10 to 20 ml 53SUNIL KUMAR.P
- 54. • Contents – Factorviii- 80 to 150 units/bag Fibrinogen- 150 to 250 mg/ bag Factor xiii- 20 to 30 % of whole blood Von wilebrand factor- 40 to 70 % of whole blood • Transfusion criteria : ABO compatibility not required Must be infused within 6 hours of thawing 54SUNIL KUMAR.P
- 55. INDICATIONS • Quantitative andqualitative fibrinogen deficiency : DIC • Correction of factor viii deficiency Haemophilia A • Von wilebrand disease • Factor XIII deficiency • Uremic coagulopathy 55SUNIL KUMAR.P
- 56. CRYOPRECIPITATE-POOR-PLASMA • It isa by-product of cryoprecipitate preparation. • It lacks labile clotting factors V and Viii and fibrinogen. • It contains adequate levels of the stable clotting factors II, VII, IX and X. • It is frozen and stored at -20°C or lower temperature for 5 years Indications • Burn 56SUNIL KUMAR.P
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- 58. ALBUMIN • Prepared froma large pool of blood reconstituted in isotonic electrolyte solution • 96% albumin and 4% globulin and other proteins Available as a 5% or 25% solutions • Half life : 6 hours • Used for hypovolemia or hypoproteinemia • Can be given without ABO Compatibility Test 58SUNIL KUMAR.P
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