--—@ YOUR POINT-BY-POINT GUIDE TO CLINICAL LABORATORY SCIENCE REVIEW aie p a se snore Pate ror ie i pct (CP)What's I. Introduction to Blood Banking and Transfusion Medicine “ Significance of Blood Banking and Transfusion Medicine “ History of Transfusion Medicine “ Roles of Medical Technologist in Transfusion Medicine Il. Who should receive blood? “ Indications and Contraindications of Blood Components itl. What should be transfused to the patient? “- Component preparation “ Contents of Blood components IV. Where do we get the blood for transfusion? “ Sources of Blood for Transfusion: Allogeneic Donation, Autologous Donation and Apheresis “ Donor Screening and Requirements V. When do we transfuse blood? “ Pretransfusion Compatibility Testing =" ABO Typing » Rh Typing » Antibody Screening - Major & Minor Blood Group Systems » Antibody Identification # Additional Techniques for Antibody Identification - Inhibition, Enzyme treatment, Adsorption, Elution & Antibody Titer Determination # Crossmatching VI. How do we transfuse blood? “ General Guidelines during Transfusion VII. Why is Transfusion Medicine important? “ Transfusion Reactions Autoimmune hemolytic anemia v “ Hemolytic Disease of the Newborn aqReferenced Modern Blood Banking & Transfusion Practices by Denise Harmening 5™ ed Fundamentals of Immunohematology: Theory and technique by Mary Louise Turgeon Henry’s Clinical Diagnosis and Management by Laboratory Methods 18" and 22™ ed Alba’s Medical Technology Board Examination Review — Volume 1, 12" ed PER Handbook eS “To read without reflecting is like eating without digesting” - E. BurkeBLOOD BANKING AND TRANSFUSION MEDICINE BLOOD BANKING - encompasses activities, procedures and tests done to ensure blood for transfusion is properly collected, preserved, stored and dispensed for later use in blood transfusion. TRANSFUSION MEDICINE - is a multidisciplinary branch of medicine that is concerned with the transfusion of blood and blood components, including proper selection and utilization of blood components as well as removal of blood or blood components in the treatment or prevention of disease. FOOD AND DRUG ADMINISTRATION (FDA) - governing body for blood bank inspections, inspects blood banks EVERY YEAR. Blood is regulated both as a BIOLOGIC and as a DRUG. MEDICINE 1492 - Transfusion for POPE INNOCENT VII from 3 human donors » 1892 - Successful transfusion by JAMES BLUNDELL OF ENGLAND to a woman suffering from postpartum hemorrhage » 1901 - Discovery of the ABO groups by KARL LANDSTEINER (author of “Specificity of Serological Reactions”) » Descatello and Sturli defined the fourth group AB > 1941 - Dr Charles Drew developed techniques on blood transfusion and blood preservation during world war II » 1943 -Loutit and Mollison of England introduced the use of ACD (acid citrate dextrose) as blood preservative Blood is botha BIOLOGIC anda DRUG » 1985 - Dr. Yves Lapierre developed gel test in Lyon, France SE hee SS Se Ne Oe Ee | eee “UNC S OF TECHNOLOGIST BLOO NKS « Recruiting blood donors = Collecting and storing whole blood or components from volunteer, paid or autologous donors * Typing, screening and preparing patient and donor blood for transfusion = Detecting and identifying unexpected antibodies in potential blood recipients or pregnant women « Establishing a database of HIV positive or questionable donors = Processing and dispensing of blood components * Performing paternity testing = Conducting tissue typing prior to organ transplantation = See. eee EA ee eee, ee, ee ee eee CONSIDERATIONS IN BLOOD BANKING AND TRANSFUSION MEDICINE: - Pre-transfusion - During the Transfusion - Post Transfusion ee ee, ee” eee ee eee ee ee eeeWHO SHOULD RECEIVE BLOOD? Transfusion Medicine was created because there are people who are in need of blood. The type of blood component to be administered to the patient depends on the need of the patient. BLOOD PRODUCT WholeBlood PURPOSE /MAJOR INDICATIONS To restore blood volume and oxygen-carrying capacity Symptomatic anemia* with large volume deficit** Packed Red Blood Cells err Washed Red Blood Cell Leuko-reduced / Leuko-poor components Neocyte-enriched red blood cells Frozen/ deglycerolized red blood cells Irradiated blood components Random Donor Platelets To restore oxygen-carrying capacity Symptomatic anemia* ~ To remove plasma proteins BO i Symptomatic anemia on patients with severe allergic or anaphylactic conditions To minimize exposure of patient to donor leukocytes _ Prevents febrile non-hemolytic transfusion reaction and non- ' . 2p : cardiogenic pulmdnary ‘edema (aka transfusion related acute lung injury) caused by antibodies against WBCs, to reduce CMV transmission ~ To prolong time between transfusions in chronically transfusion-dependent patients (e.g. sickle cell anemia or thalassemia) Prevention of non-hemolytic febrile transfusion reactions, plasma allergies or exposure to | cytomegalovirus « especially in patients with rare phenotype; autologous donation ~ To inactivate the replicative machinery of the donor leukocyte To prevent graft-vs-host-disease, for newborns and patients Bleeding*** due to quantitative or qualitative platelet defect Single Donor Platelets “if you think you are beaten, you are. If you think you dare not, you don't. If you'd like to win but think you can't, it’s almost certain you won 't” Bleeding*** due to quantitative or qualitative platelet defect, revention of HLA alloimmunization BLOOD BANKING & $i): ba'O 0) Bilis inCHECKPOINT! Blood Banking and Transfusion Medicine by Jude Me Fresh Frozen Plasma Single donor plasma Cryoprecipitate Granulocyte Concentrates Factor Concentrates Prothrombin complex Immune Serum Globulin (ISG) Volume Expanders Albumin (5%) NOTES: » *Symptomatic anemia refers to a hemoglobin level of <7 g/dL » **Average blood volume o \Male = 5-7L | o \Females = 4-6L | Vv sunt WO [PARC Hct CME | Fa tet Sela ~~ Patients with congenital hypogammaglobulinemia (given ***Platelet concentrates are usually given to actively bleeding patients who are thrombocytopenic (less than 50,000/pL) due to decreased production or decreased function A unit of whole blood or packed RBCs in an adult should increase the hematocrit level by 3% or the hemoglobin level by 1.0-1.5g/dL To replace plasma proteins and all coagulation factors except platelets in patient who are bleeding, or at risk of bleeding; multiple coagulation factor deficiency To treat hypovolemic shock or severe burns Treatment of patients with von Willebrand's disease, hilia A (factor VIII defici } or] bri ; who are bleeding, or at risk of bleeding; disseminated intravascular coagulation To elevate the leukocyte count in patients with neutropenia or granulocyte dysfunction who are suffering from, or at risk of, a life-threatening infection; Neutropenia with infection unresponsive to antibiotics Specific coagulation factor deficiencies Treatment of patients with a deficiency of factors I, VII, EX (hemophilia B) or X, who are bleeding or at risk of bleeding monthly) Patients with hemorrhagic shock and burn patients To increase blood volume while maintaining colloidal oncotic pressure Achetly beeang P\t Cconce votes Thro mh ey f Pen Z sc Ooo [ok BLOOD BANKING & By i?:): babu Bula ie i.WHAT SHOULD BE TRANSFUSED? ae C There are times when patients would not need the whole blood as a whole. There are cases where one has to receive only a part or a selected component of the blood. Component preparation is therefore, necessary. To ensure that the recipient of the blood benefits from the transfusion, blood and blood components must be checked for quality, as well for their contents. 00 O 1. Approved anticoagulant/preservatives used in blood bags contain: * Citrate - binds ionized calcium; serves as an anticoagulant , « Adenine - increases ADP levels, thereby driving glycolysis toward synthesis of ATP =. Glucose - serves as food for the cells. _ = Phosphate - serves as source of 2,3 diphosphoglycerate (2, 3 DPG) Which promotes oxygen release to tissues a 2. Anticoagulant Preservatives: Heparin 2 days (used only for priming heart-lung machines) ACD (Acid-Citrate-Dextrose) 21 days" - GPD (Citrate-Phosphate-Dextrose) 21 days CP2D (Citrate-Phosphate-Double Dextrose) 21 days CPDA-1 (citrate-Phosphate-Dextrose-Adenine) 35 days 3. Additive Solutions are preserving solutions that are added to RBCs after removal of the plasma with/without platelets. They contain saline, adenine, glucose and mannitol (SAGM). Mannitol acts as a stabilizing agent. Units with additive solution are stored up to'42 days. 4, Rejuvenating Solutions are used to restore ATP and 2, 3 DPG levels. They often contain phosphate, inosine, glucose, pyruvate and adenine (PIGPA, sometimes PIPA). RBCs stored in the liquid state can be rejuvenated at outdate or up ta.3 days after outdate and cryopreserved. 5. Atleast 70% of the red blood cells must remain viable at the end of the permitted storage period. 6. 75% of cells that have been transfused should remain viable for 24 hours. This is known as post transfusion viability. (Harmening) 7. Shipment of blood is maintained at a temperature of [1-10°O lice or other cooling devices should not physically be in contact with the blood unit to prevent hemolysis. PTL Ey Oy etic m TRANSFUSION MEDICINE "A pessimist is a person who, regardless of the present, is disappointed in the future” | 5CHECKPOINT! Blood Banking and Transfusion Medicine by Jude ee WHAT SHOULD BE TRANSFUSED? There are times when patients would not need the whole blood as a whole. There are cases where one has to receive only a part or a selected component of the blood. Component preparation is therefore, necessary. To ensure that the recipient of the blood benefits from the transfusion, blood and blood components must be checked for quality, as well for their contents. OD TLON 1. Approved anticoagulant/preservatives used in blood bags contain: = Citrate — binds ionized calcium; serves as an anticoagulant , Adenine - increases ADP levels, thereby driving glycolysis toward synthesis of ATP * Glucose - serves as food for the cells- | * Phosphate - serves as source of 2,3 diphosphoglycerate (2, 3 DPG) which promotes oxygen release to tissues — 2. Anticoagulant Preservatives: Heparin 2 days (used only for priming heart- -lung machines) ACD (Acid-Citrate-Dextrose) __ 21 days "CPD (Citrate-Phosphate-Dextrose) | ‘2idays CP2D (Citrate-Phosphate-Double Dextrose) 21 days i CPDA-1 (citrate- Phosphate- -Dextrose -Adenine) 35 days - 3. Additive Solutions are preserving 9 solutions that are added to RBCs after removal of the plasma with/without platelets. They contain saline, adenine, glucose and mannitol (SAGM). Mannitol acts as a stabilizing agent. Units with additive solution are stored up to 42 days. 4. Rejuvenating Solutions are used to restore ATP and 2, 3 DPG levels. They often contain phosphate, inosine, glucose, pyruvate and adenine (PIGPA, sometimes PIPA). RBCs stored in the liquid state can be rejuvenated at outdate or up to 3 days after outdate and cryopreserved. 5. Atleast 70% of the red blood cells must remain viable at the end of the permitted storage period. 6. 75% of cells that have been transfused should remain viable for 24 hours. This is known as post transfusion viability. (Harmening) 7. Shipment of blood is maintained at a temperature of 1-10°C Ice or other cooling devices should not physically be in contact with the blood unit to prevent hemolysis. "A pessimist is a person who, regardless of the present, is disappointed in the future” BLOOD BANKING & Bi): a/b00 ButiCHECKPOINT! Blood Banking and Transfusion Medicine by Jude 8. All units of blood should be inspected daily and prior to issue. Contamination should be suspected if the red cell mass is purple, hemolyzed, clotted or if plasma is very cloudy or discolored 3. Storage Lesions - biochemical changes in stored blood that can lead to decreased RBC viability * Decrease pH « Increased plasma hemoglobin * Decrease in glucose * Increased plasma potassium ® Decrease in ATI * Increased plasma ammonium . Decreased 2,3 DPG # Increase in lactic acid = Decreased plasma sodium 10. RBC Substitute or Blood Substitutes Hemoglobin-based oxygen carriers — this includes: = stroma-free hemoglobin ® polymerized hemoglobin * liposome-encapsulated hemoglobin - Perfluorochemicals (PFCs) UTI PONENT PREPARATION WHOLE BLOOD Packed — —— > Platelet-Rich Plasma RBC 1 Platelet rieme Cryoprecipitate 1. Packed RBCs * Can be prepared by centrifugation from: - Heavy spin (5000 x g for 5S minutes) when temperature is set at 4°C and if the whole blood used is not intended for platelet concentrate preparation; or = Light spin (3200 x g for 2-3 minutes; or 2000 x g for 3 minutes) when temperature is at 22°C and if the whole blood is to be used for platelet concentrate preparation * Unit of packed red blood cell should have enough plasma to yield a hematocrit value of not more than 80% Cryosupernate ie A v oa] fa rr eS £ io 2 wi : i ¥ : A 0 ° ol faCHECKPOINT! Blood Banking and Transfusion Medicine by Jude 2. Platelet Concentrate Can be prepared: “ From whole blood through centrifugation Platelet unit derived from whole blood is known as RDP (random donor platelet) Whole blood to be used for preparation of RDP must be processed within 6-8 hours after collection (6 hours if ACD; 8 hours for CPD, CP2D, CPDA1) Whole blood to be used for RDP preparation must be maintained at 20-24°C Unit should contain at least 5.5 x 107° platelets Unit must contain sufficient plasma to maintain a pH of greater than or equal to 6.2 (pH of 6.2 in Harmening 5" ed; 6.0 in Turgeon) Platelets units from individual whole blood can be pooled in batches of 6-8 units (or 6-10 in Turgeon) prior to transfusion. One RDP unit can increase the blood platelet count by 5,000 to 10,000 ina typical 70-g human. Steps: (1) Centrifuge whole blood using light spin (3200 x g for 2-3 minutes) to separate the red blood cells from the platelet-rich plasma. (2) Centrifuge the platelet-rich plasma using a heavy spin (3600 RPM for 5 minutes) to separate the platelets from the plasma. The plasma separated can be used to prepare fresh frozen plasma (FFP) or PF24 (plasma frozen within 24 hours) (3) Approximately 5OmL of platelets should be left after the removal of the platelet poor plasma. (4) Allow the platelet concentrate to remain undisturbed for 1-2 hours at 20-24°C. This is to enhance platelet disaggregation. Resuspend by gentle manipulation. (5) Platelets are stored at 20-24°C for 5 days in constant agitation. Agitation is important to prevent aggregation and to promote gas exchange (Rudmann). Platelet rotators used are either: e 1rpm elliptical rotator; or e 2-6rpmcircular rotators Light spin, followed by hard spin - From plateletpheresis Platelet unit obtained from plateletpheresis is known as SDP (single donor platelet) Unit should contain at least 3.0 x 10'' platelets per unit, which is equivalent to 6-10 units of random donor platelet units A unit of SDP can increase the blood platelet count by 30,000 to 60,000/ul Platelet count of the donor for plateletpheresis should be higher than 150,000/ul BLOOD BANKING & By Pita bulla *PLATELET TRANSFUSION EFFECTIVE? Calculating the Corrected Count Increment and Percent Recovery PURPOSE: Corrected Count Increment serves to assess the effectiveness of the transfusion of platelets SAMPLE PROBLEM: A patient with 10,000/uL platelet count has a body surface area (BSA) of 1.3m?. Six units of platelets are given. The one-hour post transfusion platelet count is 50,000. What is the patient's corrected increment count? Is the platelet transfusion effective? FORMULA TO BE USED: lL (post-transfusion pit count - pretransfusion pit count) x BSA number of units transfused x number of pits per unit SOLUTION: _ (50,000 /ul - 10,000 /uL) x 1.3 _ 6 units x 0.55/unit = 15,758/uL Note: 0.55 is a constant CONCLUSION: The patient has an increment of 15,758/uL. An increment of >10,000/uL is a good increment and indicates that the patient is not refractory of platelets. An increment less than 5,000/uL indicates platelet refractoriness. NOTES: = Platelet refractoriness is defined as the failure to yield an increase in recipient's platelet count on transfusion of suitably preserved platelets. The most common cause of platelet refractoriness is HLA alloimmunization. * HLA alloimmunization may result from prior exposure to donor blood components. When a recipient had previously and repeatedly been transfused by platelets from different donors, he or she may develop antibodies against the platelet and HLA antigens. These antibodies bind and destroy platelets, making platelet transfusions ineffective. = Transfusion of multiple platelet concentrates from different donors causes the formation of multiple antibodies to platelet antigens = For patients who have platelet refractoriness, the component of choice is an HLA-compatible single donor platelets. = To prevent formation of multiple antibodies to platelets, it is best to transfuse platelet units collected via plateletpheresis. « Another method that can be used to check the effectiveness of platelet transfusion is using the Percent Recovery. The expected recovery of platelets is\60% at 1 hour and 40% at 24 hours. Formula for the percent recovery is: _ (posttransfusion plt ct _- retransfusion pit ct) x blood volume ~ aproximate total number of platelets infused x 2/3 bd g M = kel = z D m m a Z f a % a oa Q 3 3 psd fa 8CHECKPOINT! Blood Banking and Transfusion Medicine by Jude 3. Plasma component = Plasma collected from platelet concentrate preparation can either be prepared as : “ Fresh Frozen Plasma (FFP) If the plasma unit has been prepared within 6 hours after collection (if ACD is used) or within 8 hours after collection (if CP2D, CPD or CPDA1 is used) “- Plasma frozen within 24 hours (PF24) - If the plasma unit has been prepared within 8-24 hours after collection « FFP is initially stored horizontally, but once frozen, should be shifted to vertical position * FFP is stored at -18°C for 1 year, or at -65°C for 7 years = FFP unit must be thawed before transfusion. Unit must be placed inside a plastic bag to prevent contamination. FFP should be thawed at 30-37°C with agitation in a water bath. "Once thawing is complete, the product should store within 1-6°C and should be transfused within 24 hours. s weed - 4. Cryoprecipitate = * Cryoprecipitate units are prepared from FFP e Steps: (1) Allow the FFP to thaw: “ Thaw slowly in the refrigerator at 1-6°C for about 14-16 hours; or “ Thaw using a circulating cryoprecipitate thaw bath (4°C water bath) for 4 hours (2) Endpoint of thawing is when plasma becomes slushy (3) Centrifuge using hard spin to separate the supernatant plasma from the precipitate. Leave approximately 10-15mL of plasma on the precipitate. The precipitate is known as the cryoprecipitate. (4) Freeze the cryoprecipitate within 1 hour from the time the plasma reached the “slushy stage” (5) The supernatant fluid can be refrozen at -18°C and labeled as “plasma cryoprecipita reduced” (also known as cryosupernatant/cryo-poor plasma, cryoprecipitate depleted) Cryoprecipitate should be thawed before issuing. Cryoprecipitate units are thawed using a 37°C water bath for up to 15 minutes. The unit should be used within 6 hours after thawing. Thawed units must be kept at room temperature until transfusion. « If several bags of cryoprecipitate are pooled, it should be used within 4 hours of pooling. Thawed units may NOT be refrozen. * Cryoprecipitate contains: factor VII, fibrinogen, factor XI, von Willebrand factor and fibronectin —_—... * Cryoprecipitate combined with thrombin results in the formation of fibrin glue. which is used to control bleeding during surgeries. * Cryosupernatant contains a variable amount of fibrinogen and a full complement of other coagulation factors apart from factor VIII and factor XIII. This component is occasionally required for the treatment of thrombotic thrombocytopenic purpura, which is resistant to exchange transfusion with large volumes of fresh frozen plasma. Cryosupernatant is devoid of FVIII/vWE multimers which are thought to be deposited on endothelial walls. Cryosupernatants can also be used as starting material for the fibrinogen content of fibrin glue. (from Modern Transfusion Medicine by Derwood Pamphilon ) BLOOD BANHING & 83) bab BulletCHECKPOINT! Blood Banking and Transfusion Medicine by Jude HOW MANY UNITS O CRYOPRECIPITATE SHOULD BE TRANSFUSED? Calculating the # of Cryoprecipitate Units Required in Factor Vill deficient patients SAMPLE PROBLEM: If the desired level of factor VIII activity is 70% and the patient's plasma volume is 2,500mL. How many single bags of cryoprecipitate units should be transfused? FORMULA TO BE USED: _ desired level of FVIIlin% x patient's plasma volume in mL 80 SOLUTION: 0.7 2,500mL = ox 6 22 bags 80 CONCLUSION: 22 single bags of cryoprecipitate are needed to achieve a factor VIII activity of 70% NOTES: Fuy= EO The constant “80” in the denominator of the formula represents the average number of units of factor VIII per single unpooled bag of cryoprecipitate. If several bags are pooled into one unit, the factor to be used in the denominator should be: 80 x number of original bags = Incases where the patient’s plasma volume is not given, plasma volume can be calculated using the following formula: PLASMA VOLUME = blood volumeinmlL x (1.0 - Hct) = From the above equation, “blood volume in mL” can be calculated using: BLOOD VOLUME = weightinkg x 70mL/kg TY) EL sce TRANSFUSION MEDICINE 3t o: hi = CHECKPOINT! Blood Banking and Transfusion Medicine by Jude SPECIAL BLOOD COMPONENTS 1. Whole blood Modified = Whole blood can be modified by having 50mL of plasma removed in the preparation of platelets, or 10-15mL of plasma removed in the preparation of cryoprecipitate 2. Irradiated Components " Irradiation is performed in order to inactivate the lymphocytes, which can cause Graft-vs- Host disease (GvHD). « Irradiation prevents GvHD but not febrile transfusion reactions " Irradiation is often done when the recipient of the blood is immunosuppressed or immunocompromised and donations to immunocompetent first-degree relatives. = Most commonly used method of irradiating blood components is by the use of a self-contained commercially available cesium!3? source. 3. Leuko-poor or Leuko-reduced components « RBC units and platelet units can undergo leuko-reduction in order to reduce the number of leukocytes « Leuko-poor units are intended to prevent febrile transfusion reactions caused by WBCs or WBC products. WBC products (also known as biologic response modifiers or BRMs) are substances which may be produced by WBCs during storage of blood. Examples of these substances are the cytokines and complement fragments. « Methods to reduce the number of leukocytes include: “ Pre-storage leuko-reduction - WBCs are removed prior to storage - Can prevent febrile transfusion reactions caused by both WBCs and BRMs. Removal of WECs before the storage can prevent the production of BRMs during storage. “ Post-storage leuko-reduction - WBCs are removed prior to issuing the unit - Can prevent febrile transfusion reactions caused by WBCs but not the BRMS since the BRMs have accumulated during the storage. Filters can remove WBCs but not the BRMs « Principles of Leuko-reduction method: “ Filtration v screen filters and micropore screen filters ¥ nylon and microaggregate filters ¥ adherence filters inverted centrifugation glucose sedimentation saline washing double centrifugation saline dilution and centrifugation G@°grtgs9s hs) BF to et ieee TRANSFUSION MEDICINECHECKPOINT! Blood Banking and Transfusion Medicine by Jude 4. Frozen & Deglycerolized Red blood cells Frozen blood are used for storage of rare blood units, as a supply of blood for autologous transfusions, or as supply of red cells having little or no plasma proteins, leukocytes or platelets Cryoprotective agents (cryoprotectants) used in freezing RBCs a. Penetrating agent - Examples include glycerol, dimethylsulfoxide (DMSO) - Small molecules; enters the cells and prevents cell dehydration as the ice forms b. Non penetrating agent - Examples include hydroxyethylstarch, glucose and polyvinylpyrrolidone (PVP) - Large molecules; does not enter the cells; forms a shell around the cells, thereby preventing loss of water Methods of Freezing Red Blood Cells a. High Glycerol Method (40% w/Vv final concentration) - Slow uncontrolled freezing; most commonly used - Stored at -80°C using a mechanical freezer b. Low Glycerol Method (20% w/v final concentration) - Rapid, more controlled freezing - Stored at-120°C using liquid nitrogen freezers c. Agglomeration - Employs a deglycerolization technique in which, in the presence of a low ionic strength solution, the cells “agglomerate”, forming large clumps that sink to the bottom of the bag - The supernatant can be removed and a wash procedure can be instituted 5. Washed RBCs Washed red blood cells are red blood cells remaining after washing with saline solution to remove the plasma Prepared by washing packed red cells by multiple batch processing through centrifugation and decanting the supernatant saline Washed RBCs must be used within 24 hours if prepared in an open system 6. RBC Aliquot Aliquots are most often transfused during the neonatal period or infants younger than 4 months of age. Be Transfusion for neonates require only small volume of RBCs (10-25mL)) It can be prepared through closed system using a multiple pack system or a quad pack system, or through an open system Aliquots from closed system retain the original outdate of the primary bag. Aliquots prepared through open system must be used within 24 hours BLOOD BANKING & 83}: ba'b i Bulla isCHECKPOINT! Blood Banking and Transfusion Medicine by Jude “ s 7. Neocyte-enriched red blood cells =" Neocytes are young red blood cells, given to patients with thalassemia or chronic conditions who are liable to get transfusion-induced hemochromatosis * Each mL of RBC contains about 1mg of iron which theoretically can be deposited in tissues and cause hemosiderosis. If neocytes with an average 90-day life span are transfused instead of conventional RBC with an average 60-day lifespan, RBC transfusion requirements and the chance of inducing hemosiderosis may be reduced " Neocytes are larger and less dense red cells and can therefore be harvested by collecting the first half of the unit during processing 8. Granulocyte concentrate = Granulocyte concentrates are given to patients with severe neutropenia who have a life- threatening systemic infection uncontrolled by antibiotics. = Granulocyte concentrates are usually prepared by leukopheresis and stored at 20-24°C for up to 24 hours. 9. Synthetic Volume Expanders * Synthetic volume expanders are used as plasma substitute for volume expansion and colloid replacement CHARACTERISTICS — Intravascular retention Poor —_ Good Peripheral edema Common Possible Pulmonary edema - Possible OS Possible a Easily excreted Yes No “Allergicreaction = + Absent = © Rare Cost Inexpensive Expensive Examples Ringer’s lactate solution Albumin (sodium, chloride, calcium, Dextran potassium and lactate) Hydroxyethyl starch 7.5% normal saline * Dextran is a plasma expander that may be used as a substitute for plasma; can be usedto treat shock by increasing blood volume. Rouleaux may be observed in the recipient’s serum or plasma. 10. Normal Serum Albumin (NSA) and Plasma Protein Fraction (PPF) = NSA and PPF are prepared from salvaged plasma, pooled and fractionated by cold alcohol process (Cohn Ethanol F ractionation) then treated with heat inactivation at 60°C for 10 hours (pasteurization). PPF has fewer purification steps compared to NSA preparation. » The heating process inactivates all the coagulation factors « NSAcontains 96% albumin.and 4% globulin. PPF contains 83% albumin and 17% globulins = NSA and PPF are indicated in patients who are hypovolemic and hypoproteinemic, and in patients with shock or burns «= NSAand PPF are stored for 5 years at 2-10°C BLOOD BANKING & Bi’: baby Bula ieCHECKPOINT! Blood Banking and Transfusion Medicine by Jude 11, Immune Serum Globulin (ISG) ISG is a concentrate of plasma globulins in an aqueous solution * ISG can be prepared from pooled plasma by cold ethanol fractionation Preparations are in the form of intravenous or intramuscular solutions ISG are indicated in patients with immunodeficiency disease 12. Factor concentrates Factor VIII, 1X, XII concentrates are lyophilized products prepared from pooled plasma using Cohn ethanol fractionation method, followed by lyophilization in order to prevent transmission of viruses Methods used to eliminate viral contamination include: “= Pasteurization - involves addition of low-molecular weight stabilizers suchas albumin, sucrose or glycine are added to prevent denaturation. The product is then heated to 60°C for 10 hours. The stabilizers are removed and the product is then lyophilized Solvent and detergent treatment - involves the use of solvent [ethyl ether and tri(n- butyl) phosphate] and detergent (sodium cholate and Tween 80) to disrupt viral coat membrane. The solvent-detergent is then removed and final product is in lyophilized form Monoclonal purification - involves immunoaffinity chromatography to positively select out of pooled plasma the coagulation factor of interest “ Ultraviolet (UV) irradiation * Factor VIII concentrates are used for treatment of classic hemophilia and hemophilia A and in persons deficient in factor VIII. Other forms of factor VIII include porcine factor VIII (xenographic form) and recombinant factor VIII. * Factor IX products are used for treatment of hemophilia B or factor IX deficiency and is available in 3 forms: “ Prothrombin complex - contains vitamin K-dependent factors (II, VII, 1X, X). This is prepared from large volume of pooled plasma by absorbing out the factors using barium sulfate or aluminum hydroxide, followed by lyophilization “ Factor IX concentrates - developed by monoclonal antibody purification “ Recombinant factor IX - produced in a Chinese hamster ovary cell line BLOOD BANKING & By \?:)\b) a'}0 0) Bull io) BaCHECKPOINT! Blood Banking and Transfusion Medicine by Jude /iag\\\ | HOW MANY Si UNITS OF FACTOR VIII Jy SHOULD BE TRANSFUSED? Calculating the # of Factor VIII units required SAMPLE PROBLEM: A 70-kg hemophiliac patient with a hematocrit level of 30% has an initial factor VIII level of 4% (4 units/dL or 0.04 units per mL). How many units of factor VIII concentrate should be given to raise his factor VIII level to 50%? FORMULA TO BE USED: Step #1 BLOOD VOL = weight in kg x 70mL/kg . * Step #2. PLASMA VOL = blood volume in mL x (1.0 - Het) % UNITS OF FACTOR VIII REQUIRED = ~. (desired factor VIII in units per mL — initial factor VIII in units per mL) Step #3 : plasma volume in mL vs SOLUTION: Step #1: Blood volume = 70kg x 70mL/kg = 4,900mL Step #2: Plasma volume = 4,900mL x (1.0 - 0.30) = 3430mL Step #3: Units of Factor VIII required = (0.50 - 0.04) x 3430 = 1578 units CONCLUSION: In order to raise the factor VIII level of the patient, 1578 units of Factor VIII should be transfused. rm BLOOD BANKING & Bi Vii biab ti BiaCHECKPOINT! Blood Banking and Transfusion Medicine by Jude ep pas — T Ks oi Avy P = & CONTENTS OF BLOOD AND BLOOD COMPONENTS (Harmening, 5" ed) BLOOD COMPONENTS STANDARDS & QUALITY STORAGE & SHELF LIFE CONTROL Whole blood Hct approx.. 40% 1- C ACD, CPD or CP2D = 21days —_ CPDA1 = 35 days Packed red blood cells Hct $80% ; - 1-6 degC a ACD, CPD or CP2D = 21days CPDA1 = 35 days ee Open Sys. = 24 hours Frozenredbloodcells © 80% RBC recovery a _ -65degC 7 <1% glycerol 10 years <300mg Hgb Washed red blood cells Hct 70-80% 1-6 degC __ 24 hours ; Irradiated¢€omponents | 25Gytothecenterofthe 1-6degC 0 canister Original outdate or 28 days from _ irradiation a ‘Leuko-reduced RBCs RBCs - components <5 x 106 Closed: same 285% RBC recovery Open: 24 hours Platelets Platelets pH 26.2; SD <5 x 10° 20-24degC; 5 days RD <8.3 x 105 Plateleteoncentrate | pH2>62 20-24degC ~~ (RDP) 25.5 x 101° 5 days Pooled: 4 hours "Single Donor Platelet pH262 20-24degC (SDP) >3.0 x 101 5 days Fresh Frozen Plasma Should be prepared within: -18degC = 1YR (FFP) 6 hours after collection for -65degC = 7YRS ACD 8 hours after collection for CPD, CP2D, CPDA1 Cryoprecipitate FVII:C = 80]U Frozen: Fibrinogen of at least -18degC, lyr 150mg per unit Thawed: 20-24degC 6hrs Pooled: 7 20-24degC, 4hrs Granulocytes 21.0 x 10'°PMNs/unit 20-24degC 24 hours BLOOD BANKING & By 3?:3.t) ato Co. By) o) Cot > oh -CHECKPOINT! Blood Banking and Transfusion Medicine by Jude | FVM &§ a s “ee ae WHERE DO WE GET BLOOD? To ensure that the recipient of blood component will benefit from the transfusion, blood must be taken from donors who have met certain sets of requirements. TYPES OF DONATION: A. Allogeneic donation - allogeneic denotes genetically different individual of the same species; allogeneic donation means that blood is taken from an individual of the same species as the recipient B. Autologous donation - autologous mean self; blood to given to the recipient came from the recipient himself C. Apheresis - process wherein blood is withdrawn from the donor and separated into its components - one or more of the components is retained and the remaining constituents are recombined and returned to the individual '’ DONOR SCREENING is important in order to assess: « Ifa donation of approximately 450mL of blood at this time be harmful for the donor * Ifthe blood drawn from the donor at this time could potentially transmit a disease to the recipient DONOR REQUIREMENTS: 1) Identification card (ID) for photographic identification 2) Donor Registration Information - full name, date and time of donation, address, telephone, sex, age and date of birth 3) Consent form - donor must sign a consent form giving permission to draw blood and perform required laboratory testing 4) Two types of tubes (red and lavender-stoppered) should be filled with blood of the donor, to be used for donor testing dete de de decitait' (ae TRANSFUSION MEDICINE "Do not let what you cannot do interfere with what you can do" 17, |