Immunohematology - 07 Blood Component Preparation

IMMUNOHEMATOLOGY BLOOD COMPONENT PREPARATION, INDICATION, AND THERAPY OUTLINE • Blood Component Preparation and Indication o Important Things to Consider o Blood Components o Whole Blood o Oxygen-Carrying Blood Products o Platelet Products o Plasma Products o Plasma Derivatives o Irradiated Blood • Apheresis o Hemapheresis o Therapeutic Cytaperesis o Therapeutic Plasmapheresis • BLOOD COMPONENT PREPARATION AND INDICATION • Aside from the selection of potential donors from the previous modules, It is also essential to study the collection and processing of donor units as well as preparation of needed components. A blood bank medical technologist must ensure that proper procedure is performed in component preparation to ensure the viability of the blood components needed in transfusion. • Two types of centrifugation o Light spin - short period (2-3 mins); 3200 rpm o Hard spin - long period (5 mins); 3600 rpm IMPORTANT THINGS TO CONSIDER • 3 components of centrifugated blood (EDTA anticoagulated) o Plasma (50-55%) - liquid component ▪ Stored frozen (-18 to 0C) ▪ Can be found on the upper portion of BB fridge o Buffy coat (<1-5%) - contains WBCs and platelets ▪ Stored at room temp (20-24C) with constant agitation ▪ Requires agitator to prevent platelet aggregation o Packed RBCs (40-45%) & Whole blood ▪ Stored refrigerated (1-6C) ▪ Can be stored on the lower portion of BB fridge • Shipping requirements for blood components o Direct - from one BB to the requesting institution ▪ Requesting facilities can bring their own temperature controlled container o RBCs & WBCS - transported with wet ice o Frozen components - transported with dry ice • Quality Control for BB fridge/ref o Temperature is monitored daily/regularly (twice/day) o Freezer = within -18 to 0C o Refrigerator = within 1-6C • Open system o Introduction of air & other contaminants o Transfused within 24 hours • Closed system o Expiration date depends on the anticoagulant present BLOOD COMPONENTS • Oxygen Carrying Products o Red cell concentrates o Leukocyte-poor red blood cells o Frozen-thawed red cells • Platelet Products o Platelet rich plasma (PRP) o Platelet concentrates (PC) • Plasma Products o Fresh frozen plasma (FPP) o Frozen plasma (FP) o Cryoprecipitate o Stored plasma • Plasma Derivatives o Normal serum albumin (NSA) o Immune serum globulin (ISG) o Factor VIII (Hemophilia A) I concentrate o Plasma protein fraction (PPF) o Rhogam o Factor IX (Hemophilia B) concentrate WHOLE BLOOD • Immediately obtained blood from donor • Indications : o Active Bleeding ▪ Can cause death when o Hemorrhagic shock o Exchange transfusion o Indicated when both oxygen-carrying capacity and volume expansion are required. • Shelf life : o CPD – 21 days o CPD-A1 – 35 days o CPD-AS-1, AS-3,AS-5 – 42 days o ACD – 21 days o CP2D – 21 days o Heparin – 2 days • Characteristics : o WBC and platelets no longer viable after 24 hrs of storage o Labile factors significantly decrease after 2d of storage ▪ Labile factors - temp-sensitive coagulation factors (Factor V & VIII) • Storage Temp : 1-6C • Immediate effect of one unit : Increase hematocrit by 1-3% OXYGEN-CARRYING BLOOD PRODUCTS PACKED RED BLOOD CELLS • Indication : Restore oxygen carrying capacity (anemia) • Contents : Hematocrit should be 80% or less • Storage Temp : 1-6C • Immediate effect of one unit : increase hematocrit by 3% and increase hemoglobin by 1g

LEUKOPOOR RED BLOOD CELLS • Indications : o Anemia with history of febrile reactions; o To decrease alloimmunization to WBC or HLA antigens or CMV transmission • Contents : 5x10 6 residual WBC • Shelf life : o Closed system – same with Packed RBC (depends on anticoagulant used) o Open System – 24 hours • Storage Temp : 1-6C • Means of Leukocyte Removal o Centrifugation o Washing procedures using saline or glycerol o Mechanical separation using leukoreduction filters ▪ First generation filters - 170 um ▪ Second generation filter - 20-40 um ▪ Third generation filter - 3-logarithmic filter REJUVENATED RED BLOOD CELLS • Addition of Rejuvenation solution (PIGPA) to regenerate ATP and 2,3-DPG o PIGPA - Phosphate, Inosine, Glucose, Pyruvate, Adenosine o PIPA - without glucose, currently used • Shelf life : Can be prepared 3 days after expiration date o Must be transfused within 24 hours o Eg. Expired blood on march 22 can be rejuvenated until march 25 • Storage Temp : 1-6C • For transfusion, wash properly and transfuse w/in 24 hours • Rejuvesol - the only FDA-approved rejuvenation solution WASHED RED BLOOD CELL • Indications : o Anemia with History of Febrile Reactions o Parxoysmal Norcturnal Hemoglobinuria o For patients with plasma proteins antibodies to reduced allergic reactions (for IgA-deficient pxs) • Shelf life : o Open System : 24 hours • Storage Temp : 1-6C • QC Requirement : Plasma removal DEGLYCEROLIZED RBCS • Frozen > Thawed > Deglycerolized RBCs • Indications : o Anemia o Long term storage of “rare’ uniis (AB- & Bombay) and/or autologous units • Shelf life : o Frozen – 10 years o Deglycerolized – 24 hours • Storage Temp : o Freezing ▪ -65C - High glycerol (40%) in mechanical freezer ▪ -120C - Low glycerol (20%) in a liquid nitrogen container or machine with temp of -120C ▪ -65C - using 79% glycerol with dextrose fructose and EDTA o Deglycerolizing Process - 1-6C PLATELET PRODUCTS RANDOM DONOR PLATELETS • Prepared from whole blood • Indications : o Thrombocytopenia (↓ platelets) o DIC o Platelet disorders o Bleeding • Shelf life : 3-5 days (5 days with continuous agitation) • Storage Temp : 20-24C with constant agitation o To prevent platelet aggregation • Contents : 5.5 x 10 10 platelet in 10-65 mL of plasma • Immediate effect : Increase platelet count by 5,000 - 10, 000 per unit agitator SINGLE DONOR PLATELET • Prepared from pheresis procedure • Indications : o Thrombocytopenia o For patients refractory to due to random platelet antibodies; do not accept platelet from other indiv. • Shelf life : o Closed system - 5 days with constant agitation o Open system - 24 hours • Storage Temp : 20-24C with constant agitation • Contents : 3.0 x 10 11 platelets in approx. 200 mL of plasma • Immediate effect : increase platelet count by 30,000 - 60, 000 per unit PLASMA PRODUCTS FRESH FROZEN PLASMA • Indication : o Treatment of multiple coagulation factor deficiencies caused by: ▪ Massive transfusion ▪ Trauma ▪ Liver disease ▪ Disseminated intravascular coagulation (DIC) o Also for treatment of Anti-thrombin III deficiency, ▪ Thrombotic thrombocytopenic purpura (TTP) ▪ Hemolytic uremic syndrome (HUS) • Shelf life : o Frozen = 1 year o Thawed = 24 hours • Storage Temp : o Frozen = -18C o Thawed = 1-6C • Contents : All coagulation factors; 400 mg Fibrinogen

SINGLE DONOR PLASMA • Indication : o Treatment of stable clotting factor deficiencies • Shelf life : o Liquid – 5 days beyond whol;e blood expiration o Frozen – 5 years • Storage Temp : o Liquid – 1-6C o Frozen --18C or colder CRYOPRECIPITATED ANTIHEMOPHILIC FACTOR • Indications : o Hemophilia A (Factor VIII:C deficiency) o Von Willebrand’s Disease (Factor VIII:R deficiency) o Fibrinogen deficiency (Factor I deficiency) o Factor XIII deficiency (stable clotting factor) • Shelf life : o Frozen – 1 year o Thawed – 6 hours o Pooled – 4 hours • Storage Temp : o Frozen = -18ºC or colder o Thawed = 20-24ºC • Contents : o Factor VIII:C - 80-150 IU o Factor VIII:vWF - 40-70% o Fibrinogen - 150-250 mg o Factor XIII - 20-30% GRANULOCYTE CONCENTRATE • Indications : o To correct severe neutropenia ( ↓ neutrophils) o Fever unresponsive to antibiotic therapy o Myeloid hypoplasia of the bone marrow • Shelf life : 24 hours • Storage Temp: 20-24C without agitation • Contents : x 10 10 WBC PLASMA DERIVATIVES • Plasma Derivatives - are concentrates of plasma proteins that are prepared from pools (many units) of plasma. FACTOR VIII CONCENTRATE • Shelf life : Varies on expiration date on vial • Storage Temp : 1-6 C (lyophilized) • Indication : Hemophilia A FACTOR IX CONCENTRATE • Shelf life : Varies on expiration date on vial • Storage Temp : 1-6 C (lyophilized) • Indication : Hemophilia B IMMUNE SERUM GLOBULIN • Shelf life: o Intramuscular: 3 years o Intravenous: 1 years • Indications: o Prophylactic treatment to pxs exposed to hepatitis, measles, or chicken pox o Treatment of congenital hypogammaglobulinemia NORMAL SERUM ALBUMIN • Shelf life & Storage Temp : o 3 years at 20-24C o 5 years at 1-6C • Contents : 96% albumin and 4% globulin • Indications : Plasma volume expansion, surgery, trauma, burns PLASMA PROTEIN FRACTION (PPF) • Shelf life & Storage Temp : o 3 years at 20-24C o 5 years at 1-6C • Contents : o 85% albumin o 15-20% globulin • Indication : plasma volume expansion SYNTHETIC VOLUME EXPANDERS • BSS • Ringer’s lactate • Electrolyte solution • Dextran • Hydroxyethyl starch (HES) Rho (D) Ig (RHOGAM) • Shelflife and Storage Temp : 3 years at 1-6C • Contents : o Full dose - 300 ug Anti-D o Mini Dose - 50 ug Anti-D • Indication : o Prevention of Rho (D) immunization (HDN) IRRADIATED BLOOD • Shelf life : 28 days or the normal fating period of the blood, whichever comes first (irradiation uses Cesium-137 or Cobalt-60) • Indications : o GVH reactions o BM transplant o Direct donation from a blood relative o Exchange transfusion o IUT o Transfusion for immunocompromised patients APHERESIS • “To separate or remove or to take from” • Enters the separation machine • Acid citrate dextrose - Anticoagulant HEMAPHERESIS • Type of blood donation where whole blood is withdrawn either from a donor or patient wherein after removal, separation, and retention of the desired cellular elements or plasma, the remaining products are recombined and returned to the donor or patient

HEMAPHRESIS DONOR SELECTION • Additional Donor Guidelines o At least 48 hours is the elapsed time after hemapheresis donation. o A donor must not exceed more than two times in a week or 24 times in a year unless otherwise allowed by bloodbank physician. o A donor must be tested to detect cytopenia (low blood cell count). ▪ Donor is tested for CBC o If a donor donates whole blood, at least 8 weeks must be elapsed before he can donate for pheresis. o Extracorporeal blood must not exceed 15% of the donor’s total blood volume. o If platelet pheresis is to be performed a donor must have above 150 x 10 9 /L platelet count. o Possible adverse reactions to HES, steroids and/or heparin must be determined. These substances are use in the apheresis procedures. THERAPEUTIC CYTAPHERESIS • Plateletpheresis o Equivalent to 6-10 random platelet concentrates o Contents: 3 x 10 11 platelets o Therapeutic Indications : ▪ Used to treat patients who have abnormally elevated platelet counts (plt. ct. > 1,000,000/uL) such as in cases of Polycythemia vera ( ↑ RBC) • Leukapheresis – remove WBC o HES (Hydroxyethyl starch) ▪ Sedimenting agent used for granulocyte colelction which causes RBCs to form rouleaux thus allowing WBCs to be harvested more efficiently o Corticosteroids → WBCs are mostly in the tissues (50% marginal pool), also present in blood vessels (50% circulating pool) ▪ Administered to the donors 12-24 hours before pheresis to increase the number of circulating granulocytes by pulling them from marginal pool o Therapeutic Indications of Leukapheresis: ▪ Used to treat patients with leukemia (WBC >100,000/uL) such as • Hairy cell leukemia • Acute myelogenous leukemia (AML) • Cutaneous T cell lymphoma • Lymphocytapheresis o Removal of lymphocytes o Therapeutic Indications : ▪ Means of producing immunosuppression in conditions like: • Rheumatoid arthritis (RA) • Systemic Lupus Erythematosus (SLE) • Kidney transplant rejection • Autoimmune disease • Alloimmune disease • Neocytapheresis o Transfusion of young RBCs “neocytes” o Therapeutic Indications : ▪ For young patients with certain hematologic disorders especially: Thalassemia syndrome (abnormal Hb synthesis) • Erythrocytapheresis o Considered an exchange procedure o Predetermined quantity of red cells is removed from the px and replaced with homologous blood o Therapeutic Indications : ▪ Used to treat various complications of sickle cell disease ( Priapism - can produce ulceration in sickle cell) and impending stroke ▪ Also in patients with severe parasitic infections from malaria and babesia THERAPEUTIC PLASMAPHERESIS • Aka Plasma Exchange • Replacement Fluids Used: o Normal Saline Solution (NSS) o Normal Serum Albumin (NSA) o Plasma Protein Fraction (PPF) o Fresh Frozen Plasma (FFP) • Therefore, FFP is now the recommended replacement fluid primarily during: o Plasma exchange for Thrombotic thrombocytopenic purpura (TTP) o Hemolytic uremic syndrome (HUS) • Therapeutic Indications : o To remove the offending agent in the plasma causing clinical symptoms in cases of paraproteinemia (e.g. Multiple myeloma, Waldenstrom macroglobulinemia, etc.), Familial Hypercholesterolemia, etc. o To collect rare RBC & WBC abs o Beneficial particularly in diseases that involve malfunction of the immune system (SLE, RA)