BLOOD COMPONENTS AND RECENT ADVANCES

BLOOD COMPONENTS AND RECENT ADVANCES-Dr. ANKITA DAS Moderator-Dr. NIRANJANA MURTHY

CONTENTSBLOOD CONSTITUENTSADVANTAGES OF COMPONENT THERAPYDISADVANTAGESQUIPMENTS FOR COMPONENT SEPARATIONAPHERESISCENTRIFUGATION PRINCIPLE COMPONENTSBLOOD COMPONENTSPLASMA DERIVATIVES RECENT ADVANCESBLOOD PRODUCTS ALTERNATIVES

ADVANTAGES OF BLOOD COMPONENT THERAPYMaximization of the yield of products from a single blood donation.

Ability to use the optimal products for specific diseases (thus minimizing the amount of unnecessary foreign material the recipient is exposed to).

The recipient can be treated with only those blood components that are lacking, reducing the occurrence of adverse transfusion reactions

More than one patient can be treated with blood components derived from one donation

Therapeutic support for patients with special transfusion requirements can be provided, for ex., plasma that often is not directly needed for transfusion can be used manufacturing of Factor VIII concentrate for Haemophilia A patients.

Improved quality and functional capacity of each components when varied storage conditions and shelf lives were applied

DISADVANTAGESPrimary disadvantage is encountered in treating patients with massive blood loss requiring massive transfusionsExposure to multiple donors increases the risk to infectious complications of transfusions

EQUIPMENTS FOR COMPONENT SEPARATION

Equipments- FreezerBlood bank refrigeratorRefrigerated centrifugePlasma expressorLaminar flowWeighing scaleDielectric sealer or aluminum clips and sealerWater bath or plasma defrosterPlatelet agitatorCryoprecipitate bath Sterile connecting device

Consumable items

350/450 ml double and triple CPDA bags

350/450 ml double or triple bags with CPD and additive solution(adsol/SAG-M)

Plastic over raps for blood bags.

Appropriate labels and instructions for each component.

Transfer packs and bag to bag connecter.

REFRIGERATED CENTRIFUGEDEEP FREEZERLAMINAR AIR FLOW BENCHPLATELET AGITATOR

DEEP FREEZERPLATELET AGITATOR

ELECTRONIC THERMAL SEALERWEIGHING BALANCEPLASMA EXPRESSORSTORAGE RACKS/TRAYS

CONSUMABLESDouble , triple or quadruple bags with or without AS

SPECIAL EQUIPMENTS

CELL SEPARATORGAMMA IRRADIATORREFRIGERATED WATER BATH

COMPONENT SEPARATION BY APHERESIS

APHERESISBlood is withdrawn from donor / patient, separated into components, one or more of the components is retained, and remaining constituents are recombined and returned to the individual

Haemonetics mcs plus LN 9000

Indications of Apheresis1. To collect the components for transfusion purpose-Donor Apheresis 2. Removing undesirable blood component -Therapeutic Apheresis.DonorTherapeuticTypes Cytapheresis Plasma -Plasmapheresis

Specific criteria for donor selection for plateletpheresis

Interval between two donations-at least 48 hrs; Donors should not undergo procedure > Twice a week / >24 times in a year (AABB STD)

2. If donor donates whole blood / red cells not returned / extra plasma being collected -Deferred for at least 8 wks.

3.If needed, platelets can be collected from donors irrespective of above criteria (HLA matched donors)

4. Donors who have taken aspirin containing medications within 36 hrs- deferred

Criteria for selection for LeukapheresisDonor Selection- ABO , Rh compatible. Free from infectious diseases(CMV inf.) HLA compatibility for alloimmunised patients Free from disorders :Diabetes, arthritis, gout, vasculitis, thrombocytopenia

Criteria for selection for PlasmapheresisPrerequisite- Written informed consent, Total count within normal limitIn cases of frequent donors--Sr. protein level of min.6.0 gm/dl is obligatory.Every 4 months, all lab tests reviewed, sr. protein electrophoresis , Ig(IgM/IgG) levels done.Safe to accept 600 ml of plasma every 2 wks.

Preparation of blood components

Different specific gravity of cellular components. red cell specific gravity 1.08 to 1.09. platelet specific gravity 1.03- 1.04. plasma specific gravity 1.02 1.03 due to different specific gravity of cellular components , they can be separated by centrifuging at different centrifugal force in g for different time.

Preparation of Blood ComponentsVarious components- separated by- CENTRIFUGATION- depends on:Relative centrifugal force (RCF)Duration of centrifugationRCF (g) = 0.0000118 x r x N2r- Radius of centrifuge rotorN- no. of rotations per min

CentrifugationPrinciple Sediment of blood cells depend on their size as well as the difference of their density from that of the surrounding fluid, viscosity of medium, flexibility of the cells which are temperature dependent

Component preparation and separation are based on the principle that different component of whole blood have different specific gravitiesRBC -1.08-1.09Platelets-1.03-1.04Plasma 1.02 1.03

Principle

27

WHOLE BLOODLIGHT SPINHEAVY SPINPRPPACKED RED CELLPLATELET POOR PLASMAHEAVY SPINPLATELET CONCENTERATEPPPLASMAFRACTIONATION

LIGHT SPIN-2000RPM FOR 3 MINHEAVY SPIN- 5000RPM FOR 5 MIN

28

COMPONENTSBlood componentsRed cell concentrate Leukocyte poor red cell concentrateWashed RBCsFrozen thawed deglycerolized RBCsGranulocyte concentrate

Platelet concentrateFresh frozen plasmaSingle donor plasmaCryoprecipitate

Plasma derivativesAlbuminPlasma protein fractionFactor VIII concentrateFactor IX concentrateImmune serum globulinRh immunoglobulin

RED CELL CONCENTRATE Prepared by removing 80% plasma from a unit of whole blood.

Red cells volume 250-300ml, approx 65-80% hematocrit and have same oxygen carrying capacity as whole blood and stored at 1-6 deg Celsius.

ADVANTAGES OF USING RBCs OVER WHOLE BLOOD:

-Equal oxygen carrying capacity in half the volume-significant reduction in level of isoagglutinins (anti-A , anti-B) facilitating safe transfusion of Group O cells to non-group O recipients.-significant reduction in acid level, citrate toxicity & potassium load.

Preparation Sedimentation Centrifugation

32

Blood is collected in double bag/ triple bag CPDA solution

Centrifuge at 3600rpm for 5 min(4-6degC

TransferSeal the tubing between primary and satellite bag using dielectric sealer

Bag containing RBCs is labelled as packed RBCsCentrifugation

solutionTransfer plasma into satellite bag using plasma expressor

33

Storage : 1-6 deg C

Shelf life: CPD=21 days CPDA-1=35 days CPD-AS=42 days

Quality controlHaematocrit: < or equal to 80%

Indications :restore O2 carrying capacity in symptomatic anemia(acute /chronic)Dosage effect:1 unit RBC: Hct 3% Hb 1 g

LEUKOCYTE REDUCED RBCsProducts in which WBC count in unit < 5* 10^6.Major indication for LRRC : prevention of non hemolytic febrile transfusion reaction particularly in multi-transfused patients or multiparous females.Depletion of leukocytes < 5* 10^8 has shown to prevent such reactions.Another indication is prevent alloimmunization to HLA antigens , reduce CMV transmission. Useful in bone marrow transplant , chemotherapy patients

Preparation:

CENTRIFUGATION-

it reduces WBC level only 70-80% .Now this procedure is replaced by more effective techniques

FILTRATION-microaggregate filters are polyester/plastic screen filters with pore size 20-40 micro-meter .usually give90-99% leukocyte reduction & recover most of RBC volume.Unit is centrifuged before filtration.Inverted spinUpright spin

Procedure is easy quick .

RBC unit prepared by this method suitable for patients with TRALI transfusion reaction.Filtration can be performed in laboratory or bed-side.

Blood bags with in-line filters that allow prestorage leukoreduction are now more commonly used.

Drawbacks of FILTER:expensiveReduce flow rate of blood if using bedside filtering procedure & if filtering is done in blood bank without expensive sterile connecting device.Open system24 hr expiration date on final product

WASHED RBCsRBCs are washed using isotonic saline by automated techniques.Saline washing step helps to remove plasma proteins.Products particularly good for patients with PNH or IgA deficiency pts with circulating anti-IgA.

Drawback- 24 hour expiration date because of open system.

RBC DEGLYCEROLIZEDA component prepared by thawing and washing red cells that have been stored in a cryoprotective agent for up to 10 years at temperatures-650C.

This freezing process is especially useful for rare blood storage.

Removal of glycerol is done by replacing glycerol with decreasing conc. of saline (12% 1.6%) with final wash in normal saline.

INDICATIONS-Storage of rare blood for a longer period -Storage of autologous blood CONTRAINDICATIONS-Not cost effective, alternatives leucocytes reduced RBC may be just as effective.

DISEASE TRANSMISSION- May transmit both hepatitis B & C-Decreases CMV seroconversion in susceptible individuals

PLATELET CONCENTRATERandom donor PlateletsWhole blood 1 unit

Platelet Concentrate 1 unit> 5.5 x 1010 platelets in 50 - 65 ml of plasma Single donor platelets1 DonorPlatelet concentrate

> 3 x 1011 platelets in 300 ml of plasma

Random Donor Platelet

Volume 50 65 ml

Single Donor PlateletVolume ~ 300 ml

PLATELET CONCENTRATEIndications Treatment of bleeding due toThrombocytopenia cancer patients during radiation , chemotherapy increased platelet consumption ( bleeding , platelet disorders, DIC , massive transfusion)

Dose and administration1 unit of PC Platelet 5000-10,000 / ul

Usual dose in thrombocytopenic patient is 6-10 units of PCChildren 1unit/10kg body weight

CCI= (post transfusion platelet increment / No. of platelet transfused x 10 11) x BSA

49

AdministrationShould be ABO & Rh compatible

After pooling, should be infused as soon as possible within 4 hours

Maintain WB at 20-24 deg C before and during platelet preparation.

QUALITY ASSURANCE OF PLATELET CONCENTRATESPrepared within 6 hours of blood collectionMust evaluate at least 4 platelet preparations monthly for platelet count, pH &plasma volumeStorage:20-240CThe temperature at which pH is measured should be the same as stored(220C)Temperature recorded at least every 4 hours during storageVolume:50-65ml(random donor platelet),200-300ml(apheresis)

pH:6.8-7.4Platelet count:4.5x1010/unit(450ml whole blood) 3.5x1010/unit(350ml whole blood) 3x1011/unit(apheresis platelet)75% units unit should fulfill these criteria at the end of storageResidual leucocytes: