Blood Component Preparation and Therapeutic Utilization

Prof. A. PourazarImmunohematologist and Transfusion medicine

School of Medicine, Isfahan Medical Science University

IRAN

Whole

Blood

Red Blood cells, Platelets and White Blood Cells- all suspended in Plasma constitute

Whole Blood.

Whole Blood UnitAfter centrifugation whole blood separates into the plasma and platelets on top and packed red blood cells on the bottom.

A plasma expresser is used to squeeze the plasma and pla

telets off the top and leave only the red blood cells in the original bag.

Blood Components

When whole blood is separated into it’s component parts we now have

Blood Components!!

Packed Red Blood Cells

Random donor Platelets

Plasma Product

1. Base Label: Bag comes with label indicating manufacturer and anticoagulant/ preservative.

2. Completed label: Paste ABO Group, Rh Type, Collection date, collecting facility etc. on label

See Harmening p235-236

Labeling the Bag

Blood Bag label when testing is complete.

Base label on bag at time of collection.

RBC Components

Packed RBCs• Approx. half the volume of Whole Blood

– Same RBC mass therefore same oxygen carrying capacity

• Total Volume: 250-300 ml

• Hematocrit <80% for CPDA-1 anticoagulant/ preservative.

• Expiration Date: depends on sterility and anticoagulant/preservative– CPDA 1 - 35 days closed system

– AS-1 - 42 days closed system

– Open System - 24 hours

RBC:Anticoagulant/Preservative Solutions

Purpose of RBC PreservationDesigned to prevent clotting and maintain red cell

viability and function during storage.

Anticoagulant-Preservative ContentsCitrate: anticoagulant (chelates calcium)Dextrose: ATP generation via glycolytic pathwayAdenine: Acts as a substrate for RBC synthesis of ATPSodium diphosphate: Buffer to control decrease of

pH expected from generation of lactic acid over time.

RBC Anticoagulant/Preservative Solutions

Cold Temperature • RBC components are kept at 1-6oC in a

monitored refrigerator• Inhibits bacterial growth • Slows glycolytic activity: But RBCs

continue metabolic activity during storage consuming nutrients and depleting intracellular energy sources.

RBC Component Storage Lesion

• Definition: The biochemical changes that occur during storage of RBC components.

• Changes in Red Blood Cells include:

2,3-DPG, K+, ATP Na+

• Changes in the PLASMA include:

K+, Ammonia, pH, and haemolysis

Indications for Transfusion of Packed Red Blood Cells

• Need for blood depends on patients:– Oxygen demand

– Cardiac Output– Haemoglobin concentration (Hgb and Hct)

• Each unit of transfused RBCs increases

-Hemoglobin by 1.0 g/dL

-Hct by 3%.

Indications for Transfusion of Packed Red Blood Cells…

RBC Transfusion Categories1. Iatrogenic: neonatal transfusions often

result from withdrawal of too much blood for testing.

2. Decreased Bone Marrow Production: Leukemia, aplastic anemia, etc.

3. Decreased RBC Survival: Hemolytic anemia, etc.

4. Surgical/Trauma: Excessive bleeding

Indications for Transfusion of Packed Red Blood Cells…

Fresh Red Blood Cells (<7 days old)• Neonatal transfusion. Why?• Massive Transfusion is defined as replacing the

patients entire blood volume within 24 hours. • Why would we want to transfuse fresh blood in this

situation?2,3-DPG levels drop within 10 to 15 days of collection

to levels that will NOT adequately oxygenate the tissues. Also want to avoid transfusion of excess K+, ammonia

and haemolysis.

RBC Components continued…Leukocyte reduced RBC: to a level of

<5.0 X 106 per unit• Leukocyte filtration: removes WBCs,

- see p242• Washing- removes WBCs, not efficiently,

24 hour outdate, -see p242• Freezing/Deglycerolization: Frozen RBC

unit is thawed and washed to remove both glycerol and WBCs, see p242

Indications for Transfusion of Leukocyte Reduced RBCs

• Febrile non haemolytic transfusion reactions– Patient history of febrile transfusion reactions

• HLA allo-immunization: antibody to HLA antigens

• Transfusion related acute lung injury (TRALI)• Transfusion associated Graft vs. Host disease

(TA-GvHD).• Transfusion related immune suppression:

- Reduces risk of transmission of CMV and HIV.

RBC Components continued…

Frozen, Deglycerolized RBC• Freezing RBCs in glycerol gives minimal

damage to the cell.• >10 years Shelf life, while in frozen, for

‘rare’ blood types• Free (almost) of WBCs, Platelets & Plasma• Indicated for patient with anti-IgA; Intra-uterine transfusion (as lacks WBCs)• Two methods: Page 242-243

Platelet Components

Includes Random donor platelets, Single donor platelets and Pooled Platelets.

Platelet Components Preparation

• Random Donor From a Whole Blood (WB) Unit at kept 20-24oC,

Step 1: WB centrifuged using a Soft (light) Spin for 2-3 min at 3200 rpm (to keep the platelets in the plasma).

Step 2: Separate platelet rich plasma from RBCs and using a Hard (heavy) Spin (5 min at 3600 rpm) to settle platelets.

Step 3: Express off all but 55- 65 ml of plasma and let platelets rest and resuspend at RT for 1-2 hours

• Single Donor (by apheresis)

Platelet Components

Random Donor Platelet• At least 5.5 x 1010 platelets/unit• Store at 20 to 24oC with continuous agitation• Suspended in 55-65 ml plasma• Shelf life of 5 days

Single Donor Platelet - Apheresis• At least 3.0 x 1011 platelets/unit• Store at 22 to 24oC with agitation• Suspended in 300 ml plasma (equivalent to 4-8

random donor platelets)• Shelf life of 5 days

Platelet and Plateletpheresis• Platelet Increment (desired): Patient platelet

count should increase by 5-10,000/ random donor platelet transfusion and 30-60,000/ per single donor platelet.

ABO and Rh Compatibility• Paediatric: Transfuse ABO/Rh compatible units Volume may be reduced by

• Adults:Want to use ABO compatible with adults but it is NOT necessary. Need to be careful since there is 50-60 ml of plasma per unit.

No cross match is necessary.

Transfusion of platelets is indicated when patient’s bleeding results from decreased

platelet count. Platelet ProductsPlatelet refractoriness (Poor platelet

increment)

• ITP (idiopathic thrombocytopenic purpura) and

• TTP (thrombotic thrombocytopenic purpura) – Due to antibody to platelets: don’t transfuse unless

absolutely necessary (ITP and TTP). Adding fuel to the fire.

• Splenomegaly, drugs or sepsis can also cause platelet refractoriness.

Indications for Transfusion of Platelets

• Thrombocytopenia Decreased platelet count

• Causes:Decreased Platelet production: chemo-

therapy, malignancy, etc.Increased Platelet destruction: DICDecrease by dilution: Massive transfusion

• Transfusion indicated, if associated with bleeding

Plasma Components

Fresh Frozen Plasma• Plasma expressed from Whole Blood, frozen

within 8 hours of collection for CPDA-1 anticoagulant (6 hrs for ACD).

• Frozen at -18oC: 1 year expiration date• Frozen at -65oC: 7 year expiration date• Thawed: 24 hour kept at 1-6oC• 150-250 ml total volume• Contains all clotting factors

Indications for Transfusion of Plasma Products

Fresh Frozen Plasma (FFP)• Treat multiple coagulation factor deficiencies

such as DIC, liver failure, vitamin K deficiency, or massive transfusion

• Good for factor deficiency where there is no suitable clotting factor concentrate: Factor XI deficiency

• NEED to be ABO compatible. No cross-match needed

• Contraindicated for volume expansion and protein replacement.

Plasma Substitutes• Colloid SolutionsAlbumin: 5% and 25%Plasma Protein Fraction: 5% albumin and non

albumin plasma proteins

• Crystalloid Solutions Normal saline, lactated Ringers, dextran• Indications: Volume expansion/protein replacement

without risk of transfusion transmitted viruses.

• Advantages over plasma: No disease transmission, cost, reduced risk of allergic reactions.

Cryoprecipitated Antihemophilic Factor (Cryo)

• Cold insoluble portion of Plasma• Preparation Procedure (Page 246)

– Take FFP and thaw at 1 to 6oC until it becomes slushy (14-16 hours)

– centrifuged using Hard Spin (5-7 min at 3500 rpm).

– Remove cryo-poor plasma leaving about 10-15 ml of plasma with Cryo-precipitates.

– Freeze Cryo at -18oC for 12 months

Cryoprecipitated Antihemophilic Factor (Cryo)

• Contains at least 80 units of AHF and 150-250 mg Fibrinogen

• Also contains Factor XIII and von Willebrands factor

• Must be transfused within 6 hours of thawing or within 4 hours of pooling

• Once thawed store at room temperature until transfusion

Indications for Transfusion of Cryoprecipitated AHF

• Primary use: intravenous supplementation of Factor XIII and fibrinogen

• Topical Use: Fibrin sealant (glue) in surgery

• Factor concentrates (i.e. Factor VIII) have replaced CRYO in many situations because of reduced risk of transmission of disease.

• See Table 16-4, Page 349 Harmening

Granulocyte Concentrates• Prepared by Pheresis (Chapter 17)

– Single donor component

• Should contain 1 x 1010 granulocytes/unit• 200-600 ml plasma and red cells

– Must be ABO and Crossmatch compatible with recipient

• Store at 20oC to 24oC • Shelf life is 24 hours

Indications for Transfusion of Granulocyte

Patient should meet the following conditions:1. Neutropenia: WBC count <500/mL

2. Fever for 20-48 hours, positive bacterial or fungal cultures, unresponsive to antibiotic therapy.

3. Myeloid hypoplasia

4. A reasonable chance for recovery of marrow function.

• Component should be irradiated and CMV seronegative.

Irradiated Blood Components

• Gamma irradiation of component• Purpose: Inactivate donor lymphocytes to

prevent Graft Vs. Host disease (GVHD). Only accepted method.

• Expiration date: 28 days (RBCs) from irradiation or original outdate - which ever comes first.

Indications for Transfusion of Irradiated Blood Products

1. Congenital Immunodeficiency's: SCID, Di George Syndrome

2. Hodgkin's Lymphoma

3. Bone Marrow Transplant

4. Intrauterine Transfusion to Fetus

5. Exchange transfusion: Neonate

6. Transfusion of first degree relative: Blood relative

ALL BLOOD COMPONENTS ARE ADMINISTERED THROUGH A

FILTER!

• Necessary to remove any accumulated debris that may be present in the blood component such small clots, fibrin, etc.

Autologous Blood Transfusion

• Donation from the recipient,

• Rare blood type

• Having antibody to high frequency antigen

• Having a presence of multiple CSA

• Useful in planned surgery

• Large number of units by leap-frog method

Autologus Transfusion:•Leap-Frog collection Approach•5 units collected over 5 weeks prior to surgery•Iron supplement to the patient•Hb to be monitored: Hb, 11g%; HCT, 0.34: ok •Informed consent from the patient needed •Unit’s identity: label to include patient’s signature•Stored in separate-special area

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