BB Lecture 8 - Blood and Blood Components

Manual - collection in a bag

Automated - collection on an aphaeresis machine

What are the 2 methods for collecting blood/blood components?

- Non Leuko-reduced (LR)

- LR

What are the components in whole blood?

➢ RBCs (non LR)

➢ LR-RBCs

➢ Washed (W-RBCs)

➢ Frozen / Deglycerolized (F-RBCs / Deglyc-RBCs)

What are the components in Red Blood Cells aka Packed cells?

- LR

- Non-LR

What are the components of platelets?

✓ Platelet Concentrates (Plt Conc) - made from a whole blood donation; commonly call "Random"

✓ Plateletpheresis (Pltph) - collected by an apheresis machine

✓ Cold-stored Platelets

List the types of products that stem from Platelets

➢ Fresh Frozen Plasma (FFP)

➢ Plasma Frozen within 24 hrs from collection (PF24)

➢ Cryoprecipitate (Cryo)

➢ NEW- Freeze-Dried Plasma (FDP)

List the components in Plasma

1. CPD (Citrate Phosphate Dextrose)

2. CP2D (Citrate Phosphate Double Dextrose)

3. ACD (Acid Citrate Dextrose)

4. CPDA-1 (Citrate Phosphate Dextrose Adenine)

5. Additive Solutions

➢ AS-1 (Adsol)

➢ AS-3 (Nutricel)

➢ AS-5 (Optisol)

Identify the 5 common types of anticoagulant preservatives used in the USA.

1. CPD

2. CP2D

3. ACD

4. CPDA-1

5. Additive Solutions

➢ AS-1

➢ AS-3

➢ AS-5

CPD, CP2D, ACD

Which preservatives have a 21 day expiration?

CPDA-1

Which preservative has a 35 day expiration

Additive solutions

Which preservatives have a 42 day expiration

➢ ↓ Glucose

➢ ↓ ATP (causes ↑ Hgb O2 affinity)

➢ ↓ 2,3 DPG declines rapidly after 1-2 wks (causes ↑ Hgb O2 affinity)

➢ ↓ pH

➢ ↑ K+ in supernatant

➢ ↑ free Hgb

In RBC storage, what are the biochemical changes?

➢ Irreversible shape deformity

➢ ↑ Osmotic Fragility

In RBC storage, what are the morphological changes?

➢ ↓ Glucose

➢ ↓ pH

➢ ↑ lactate dehydrogenase (LDH)

In platelet storage, what are the biochemical changes?

➢ Very fragile / fragmentation; degranulation occurs

➢ Without continuous rotation, platelets will clump together

➢ Irreversible shape deformity from discoid to spherical

➢ Decreased in-vivo recovery, survival, & function

In platelet storage, what are the morphological changes?

➢ Deterioration of heat-labile coagulation factors, i.e., Factor V & VIII

In plasma storage, what are the biochemical changes?

➢ There are no cellular elements found in plasma products, therefore, there is no morphological changes occurring.

In plasma storage, what are the morphological changes?

RBC - 120 days

Platelets ~ 10 days

WBCs ~1-3 cays

Coag Factors (half-life) ~ hours

List the life spans of each in the body

RBC -

Platelets -

WBC -

Coat Factors (half-life) -

- Truma or massive bleed

- Hypovolemia

- Increase oxygen-carrying capacity to vital organs

Whole Blood Indications

- Acute hemorrhaging:

- Volume replacement:

- Increase in what to vital organs?

❑ Volume: 400 - 500 ml

❑ Storage Temp: 1 - 6°C

❑ Expiration: ACD, CPD, or CP2D - 21 days

❑ Transport Temp: 1 - 10°C

❑ Contents: RBCs, Plasma, WBCs, & Plts

❑ Quality Control Requirements: None

Fill in the Whole Blood characteristics

❑ Volume:

❑ Storage Temp:

❑ Expiration:

❑ Transport Temp:

❑ Contents:

❑ Quality Control Requirements:

❑ Volume: 400 - 500 ml

❑ Storage Temp: 1 - 6°C

❑ Expiration: ACD, CPD, or CP2D - 21 days

❑ Transport Temp: 1 - 10°C

❑ Contents: RBCs, Plasma, WBCs, & Plts

❑ Quality Control Requirements:

➢ A & B hemagglutinin titer is < 1:256; can be lower for more conservative titer

➢ Must be labeled as "Low Titer"

Fill in: Low Titer O Whole Blood Characteristics:

❑ Volume:

❑ Storage Temp:

❑ Expiration:

❑ Transport Temp:

❑ Contents:

❑ Quality Control Requirements:

❑ Volume: 400 - 500 ml

❑ Storage Temp: 1 - 6°C

❑ Expiration: CPD or CP2D - 21 days; CPDA1 - 35 days

❑ Transport Temp: 1 - 10°C

❑ Contents: RBCs, Plasma, WBCs, & Plts

❑ Quality Control Requirements:

➢ < 5x106 WBCs & ≥ 85% RBC Recovery in at least 95% of the units tested

Fill in Leuko-Reduced (LR) Whole Blood characteristics

❑ Volume:

❑ Storage Temp:

❑ Expiration:

❑ Transport Temp:

❑ Contents:

❑ Quality Control Requirements:

1. RBC

2. RBC with an additive solution

3. Leuko-reduced RBC

What are the 3 types of RBC components?

❑ Anemia condition

❑ Less volume; helps prevent hypervolemia

❑ Increase oxygen-carrying capacity to vital organs

❑ 98% of blood transfusions is using this product

❑ Leuko-reduced RBCs are majority of the blood made / available

❑ 1 unit of RBCs raises Hgb ~ 1 gm/dL or Hct ~ 3%

What are the indications of RBCs?

❑ Volume: ≈ 250 ml (no additive sol't used)

❑ Storage Temp: 1 - 6°C

❑ Expiration: 21 days

❑ Transport Temp: 1 - 10°C

❑ Contents: RBCs, plasma, residual WBCs & Plts

❑ QC Requirements:

➢ RBC Hct ≤ 80% for non-additive RBC product

Fill in the RBC characteristics

❑ Volume:

❑ Storage Temp:

❑ Expiration:

❑ Transport Temp:

❑ Contents:

❑ QC Requirements:

❑ Volume: ≈ 300 ml

❑ Storage Temp: 1 - 6°C

❑ Expiration: 42 days

❑ Transport Temp: 1 - 10°C

❑ Contents: RBCs, plasma, residual WBCs, additive solution, & Plts

❑ QC Requirements:

➢ Must add additive solution to RBCs within 3 days from collection when temp

is maintained at 1-6°C.

➢ No Hct QC on additive RBC product as final product Hct after adding the

additive solution is 55 - 65%

Fill in the Additive-RBCs characteristics

❑ Volume:

❑ Storage Temp:

❑ Expiration:

❑ Transport Temp:

❑ Contents:

❑ QC Requirements:

- Helps reduce Febrile Transfusion Reacts and Transfusion-related Acute Lung Injury Transfusion Reactions due to lack/reduced number of WBCs and plts in the produce

- Helps reduce the recipients immune response to develop Abs against WBCs and platelets

What are the other advantages of LR-RBCs indications?

❑ Volume: 300 ml

❑ Storage Temp: 1 - 6°C

❑ Expiration: 42 days

❑ Transport Temp: 1 - 10°C

❑ Contents: RBCs & additive Sol't; essentially no WBCs or Plts

❑ Quality Control Requirements:

➢ < 5x106 WBCs & ≥ 85% RBC Recovery in at least 95% of the units tested

Fill in LR-RBCs characteristics

❑ Volume:

❑ Storage Temp:

❑ Expiration:

❑ Transport Temp:

❑ Contents:

❑ Quality Control Requirements:

➢ Removes plasma proteins (e.g., IgA, IgG, etc) during washing cycles

➢ Removes other accumulated substances present in stored blood, e.g., K+

What are the W-RBCs advantages?

❑ Volume: 180 - 200 ml

❑ Storage Temp: 1 - 6°C

❑ Expiration: 24 hrs (open system* process)

❑ Transport Temp: 1 - 10°C

❑ Contents: RBCs, Saline

❑ Quality Control Requirements:

➢ Product Hct 70-80%

Fill in the W-RBCs characteristics

❑ Volume:

❑ Storage Temp:

❑ Expiration:

❑ Transport Temp:

❑ Contents:

❑ Quality Control Requirements:

❑ Same as W-RBCs, but with other advantages:

➢ Rare phenotypes are frozen for future transfusions, e.g., hr" neg, Oh , U

neg, etc.

➢ Autologous blood

F-RBC indications and advantages

❑ Volume: 250 - 300 ml

❑ Cryoprotective Agents Used:

➢ High Glycerol (40%) - most commonly used in the USA

➢ Low Glycerol (15-20%)

❑ Limitation: must be frozen within 6 days from collection

❑ Storage Temp:

➢ High Glycerol: ≤ − 65°C in a mechanical BB ultra-low freezer (usually @ − 80°C)

➢ Low Glycerol: ≤ − 165°C in liquid nitrogen vapor OR ≤ − 196°C in liquid nitrogen in a monitored liquid nitrogen tank

❑ Expiration: 10 yrs

❑ Transport Temp:

- High Glyc - Dry Ice

- Low Glyc - Liquid N2

❑ Contents: RBCs and Cryoprotective Agent

❑ Quality Control Requirements: None

Fill in F-RBCs characteristics

❑ Volume:

❑ Cryoprotective Agents Used (2):

❑ Limitation:

❑ Storage Temp (2):

❑ Expiration:

❑ Transport Temp (2):

❑ Contents:

❑ Quality Control Requirements:

Using varying dilutions of saline with dextrose

deglycerolization

Prior to transfusion how is the removal of all the cryoprotective agent performed?

What is this removal process called?

❑ Volume: 250 ml

❑ Storage Temp: 1 - 6°C

❑ Expiration: 24 hrs (open system process)

❑ Transport Temp: 1 - 10°C

❑ Contents: RBCs & 0.9% Saline w 0.2% dextrose

❑ Quality Control Requirements:

➢ ≥ 80% RBC Recovery

➢ < 1% Residual Glycerol

Fill in the DEGLYC RBCs characteristics

❑ Volume:

❑ Storage Temp:

❑ Expiration:

❑ Transport Temp:

❑ Contents:

❑ Quality Control Requirements:

➢ Lyophilizing process affect on RBCs

➢ Drying damage affects

➢ Rehydration issues

Although it will be a new future product, what are some issues with RBC lyophilization thus far?

a whole blood collection using a 2-step differential centrifugation process, i.e., low RPM, then high RPM

What are platelet products made from?

➢ Based on the number of platelets transfused and the size of the patient (BSA) and is used to measure platelet refractoriness.

➢ Performed 10 - 60 mins post plt transfusion

➢ Normal: > 7,500 µL & above 4,500 µL at 24 hrs

➢ Abnormal: < 5,000 µL

(Plt Conc. Indications)

Correct Count Increment characteristics

bleeding, spleen sequestration, alloimmunization / plt Ab,

disease state, etc.

What are the factors that platelet transfusion responses are dependent on?

❑ Volume: 50 - 70 ml

❑ Storage Temp: 20 - 24°C with constant agitation

❑ Expirations: 5 days; 7 days with performance of bacterial detection testing; Pooled (4-6 units / pool): 4 hrs

❑ Transport Temp: 20 - 24°C

❑ Contents: Plts, WBCs, residual RBCs, Plasma

❑ Quality Control Requirements:

➢ ≥ 5.5 x 1010 Plts

➢ pH ≥ 6.2

Fill in the PLT Conc. Characteristics

❑ Volume:

❑ Storage Temp:

❑ Expirations:

❑ Transport Temp:

❑ Contents:

❑ Quality Control Requirements:

❑ Volume: 50 - 70 ml

❑ Storage Temp: 20 - 24°C with constant agitation

❑ Expiration: 5 days; Pooled: 4 hrs

❑ Transport Temp: 20 - 24°C

❑ Contents: Plts, residual RBCs, Plasma

❑ Quality Control Requirements:

➢ ≥ 5.5 x 1010 Plts

➢ pH ≥ 6.2

➢ < 8.3x105 WBCs in 95% of the units tested

Fill in LR Platelet Conc Characteristics

❑ Volume:

❑ Storage Temp:

❑ Expiration:

❑ Transport Temp:

❑ Contents:

❑ Quality Control Requirements:

- Replacement of all coagulation factors

- Massive hemorrhage: hypovolemia

= Re-establishes normal coagulation scheme in massive hemorrhage

Fresh Frozen Plasma (FFP) indications

❑ Volume: 200 - 250 ml

❑ Storage Temp: ≤ − 18°C

❑ Limitation:

➢ Immediately after collection, WB must be rapidly cooled (via ice chest containing wet ice or BB refrigerator) to 1 - 6°C

➢ Must be separated and frozen within 8 hrs from collection

❑ Expiration: Frozen - 1 yr

❑ Expiration Thaw for transfusion: thaw FFP in 37°C water bath; expiration &

storage - 24 hrs at 1 - 6 °C

❑ Transport Temp: Frozen - ≤ − 18°C; Liquid thawed FFP: 1 - 6°C

❑ Contents: Plasma, no cellular elements

❑ Quality Control Requirements: None

FFP characteristics

❑ Volume:

❑ Storage Temp:

❑ Limitation:

❑ Expiration:

❑ Expiration Thaw for transfusion:

❑ Transport Temp:

❑ Contents:

❑ Quality Control Requirements:

- Product is NOT frozen within the same time limit as FFP, but within 24 hours from collection

- used only as liquid plasma because the heat-liable coat factors are gone, but the more stable coag factors are still present

Plasma frozen within 24 hours from collection (PF24) - Indications:

❑ Volume: 200 - 250 ml

❑ Storage Temp: ≤ − 18°C

❑ Limitation:

➢ Immediately after collection, WB must be stored a 1 - 6°C within 24 hrs from collection

➢ Must be separated and frozen within 24 hrs from collection

❑ Expiration: Frozen - 1 yr

❑ Expiration Thaw for transfusion: thaw PF24 in 37°C water bath; expiration & storage - 24 hrs at 1 - 6°C

❑ Transport Temp: Frozen - ≤ − 18°C; Liquid thawed PF24: 1 - 6°C

❑ Contents: Plasma, no cellular elements

❑ Quality Control Requirements: None

PF24 Characteristics

❑ Volume:

❑ Storage Temp:

❑ Limitation:

❑ Expiration:

❑ Expiration

❑ Transport Temp:

❑ Contents:

❑ Quality Control Requirements:

❑ Thawed FFP or PF24 stored beyond the original 24 hr expiration can be converted / relabeled to the status of "thawed plasma"

❑ Volume: same as FFP & PF24

❑ Storage Temp: 1 - 6°C

❑ Expiration: 4 days after initial thaw date

❑ Transport Temp: 1 - 10°C

❑ Contents: Reduced Factor V & VII levels; adequate amounts of other factors

❑ Quality Control Requirements: None

Thawed Plasma Characteristics

❑ Thawed FFP or PF24 stored beyond the original 24 hr expiration can be converted / relabeled to the status of "thawed plasma"

❑ Volume:

❑ Storage Temp:

❑ Expiration:

❑ Transport Temp:

❑ Contents:

❑ Quality Control Requirements:

Freeze-Dried Plasma (FDP)

What type of plasma was approved for American military use only?

❑ Volume: 260 - 270 ml OR 700 ml (apheresis)

❑ Storage Temp: Room Temperature

❑ Expiration: 2 yrs

❑ Transport Temp: Room Temperature

❑ Contents: Plasma, no cellular elements

❑ Quality Control Requirements: ??? TBD*

FDP characteristics

❑ Volume:

❑ Storage Temp:

❑ Expiration:

❑ Transport Temp:

❑ Contents:

❑ Quality Control Requirements:

- Storage temperature is room temp

- Reconstitution is quick

- Maintaining inventory of AB plasma in rural hospitals

- Use in the field to increase survivability/decrease mortality

Advantages of FDP

❑ Replacement of Factor VIII or Fibrinogen or vWF, but primarily vused to replace Fibrinogen

❑ Each unit raises the fibrinogen level 5-10 mg/dL

❑ Can be used to make biologic fibrin glue in certain surgeries to stop ooze bleeding

Cryoprecipitate (Cryo) Indications

❑ Made from FFP

❑ Volume: 10 - 25 ml

❑ Storage Temp: Frozen - ≤ −18°C; for transfusing

❑ Expiration: Frozen- 1 yr;

❑ Expiration Thaw for transfusion: thaw cryo in 37°C water bath; storage & expiration -

store at 20 - 24°C for: a) 6 hrs as individual unit or b) Pooled (4 - 10 units) - 4 hrs in

"open" system

❑ Transport Temp: Same as Storage Temp

❑ Contents: Fibrinogen, Factor VIII, von Willebrand's Factor (vWF); some Plasma

❑ Quality Control Requirements: ≥ 80 IU Fac VIII; ≥ 150 mg Fibrinogen

Cryo (Individual Unit) Characteristics

❑ Made from FFP

❑ Volume:

❑ Storage Temp:

❑ Expiration:

❑ Expiration Thaw for transfusion:

❑ Transport Temp:

❑ Contents:

❑ Quality Control Requirements:

❑ Made from FFP

❑ Individual units are pooled prior to initial freezing; pooling involves usually 4 - 10

units; pooling process is using a sterile connecting device, aka a "closed system"

❑ Volume: Dependent on the number of units in the pooled unit

❑ Storage Temp: Frozen - ≤ −18°C

❑ Expiration: Frozen- 1 yr;

❑ Expiration Thaw for transfusion: thaw pooled cryo in 37°C water bath; storage & expiration - store at 20 - 24°C for 6 hrs

❑ Transport Temp: Same as Storage Temp or if thawed, 20 - 24 °C

❑ Contents: Fibrinogen, Factor VIII, von Willebrand's Factor (vWF); some Plasma

❑ Quality Control Requirements: ≥ 80 IU Fac VIII; ≥ 150 mg Fibrinogen

Cryo (Individual Unites Pooled Before Freezing) Characteristics

❑ Made from FFP

❑ Individual units are pooled prior to initial freezing; pooling involves usually 4 - 10 units; pooling process is using a sterile connecting device, aka a "closed system"

❑ Volume:

❑ Storage Temp:

❑ Expiration:

❑ Expiration Thaw for transfusion:

❑ Transport Temp:

❑ Contents:

❑ Quality Control Requirements:

Sterile Connecting Device

Device that connects 2 separate blood tubing without breaching the integrity of either container/aliquot

To select certain blood elements to be collected and returning the remaining blood to the donor

What capability does the aphaeresis machine have?

- referred to as Single donor Platelets

- are larger in volume

- have increased quantities of cellular elements

- uses less manipulation in processing the product when compared to the WB-derived components

- can collect both non Leuko-reduced products and Leuko-reduced products without the added filtration processing step

Apheresis products

❑ Pltph product

➢ Is equivalent to ≈ 4-6 Plt concs

➢ Raises patient Plt Ct after transfusion to ≈ 40,000/μL in a 70-kg adult

❑ Corrected Count Increment (CCI) is also used to determine platelet refractoriness.

❑ Platelet transfusion response dependent on a number of factors, e.g., bleeding, spleen sequestration, alloimmunization /

plt Ab, disease state, etc.

Plateletpheresis (PLTPH) Indications

❑ Volume: 200 - 300 ml

❑ Storage Temp: 20 - 24°C with constant agitation

❑ Expiration: 5 days OR 7 days (bacterial reduction)

❑ Transport Temp: 20 - 24°C

❑ Contents: Plts, WBCs, no RBCs, Plasma

❑ Quality Control Requirements:

➢ ≥ 3.0 x 10^11 Plts

➢ pH ≥ 6.2

PLTPH - Characteristics

❑ Volume:

❑ Storage Temp:

❑ Expiration:

❑ Transport Temp:

❑ Contents:

❑ Quality Control Requirements:

❑ Volume: 200 - 300 ml

❑ Storage Temp: 20 - 24°C with constant agitation

❑ Expiration: 5 days OR 7 days (bacterial reduction)

❑ Transport Temp: 20 - 24°C

❑ Contents: Plts, Plasma, no RBCs or WBCs

❑ Quality Control Requirements:

➢ ≥ 3.0 x 1011 Plts

➢ pH ≥ 6.2

➢ < 5x106 residual WBC in 95% of the units tested

Leuko Reduced (LR PLTPH) - Characteristics

❑ Volume:

❑ Storage Temp:

❑ Expiration:

❑ Transport Temp:

❑ Contents:

❑ Quality Control Requirements:

❑ Stops / controls bleeding almost "immediately"

❑ The cold temperature alters the platelet morphology (from discoid to spherical with appendages) which increases its ability to quickly adhere to the wound site / enhance their ability to participate in the hemostatic process

❑ Reduced potential of bacterial proliferation

Cold-Stored Platelet - Indications

❑ Volume: 200 - 400 ml

❑ Storage Temp: 1 - 6°C with NO agitation

❑ Expiration: 14 days

❑ Transport Temp: 1 - 10°C

❑ Contents: Plts, WBCs, no RBCs, Plasma

❑ Quality Control Requirements:

➢ Immediately after collection, product placed in 1-6C storage

➢ Pathogen-reduction process implemented within 4 hrs from collection

➢ Ensure proper storage temperature (1-6C); cannot be returned to 20-24C

➢ pH ≥ 6.2

Cold-stored Plateletpheresis - Characteristics

❑ Volume:

❑ Storage Temp:

❑ Expiration:

❑ Transport Temp:

❑ Contents:

❑ Quality Control Requirements:

RBC Apheresis/Erythrocytaapheresis

Apheresis products that are collected, can be either a single or double product, depending on the donor's pre-donation Hgb/Hct values

➢ Treatment of profound neutropenia (< 500/µL or 0.5x109 / L) in the presence of bacterial or fungal infection unresponsive to antimicrobial therapy.

➢ Transfusion of this product should continue at least daily until the infection is cured.

➢ Because most recipients of this product are severely immunosuppressed, product should be irradiated prior to transfusion to

prevent TA-GVHD.

Granulocytes Apheresis Indications

❑ Volume: 200 - 300 ml

❑ Storage Temp: 20 - 24°C with NO agitation

❑ Expiration: 24 hrs

❑ Transport Temp: 20 - 24°C

❑ Contents: Plts, WBCs, RBCs, Plasma containing HES (a sedimenting agent)

❑ Limitations:

➢ Due to large amount of RBCs present (≥ 20 ml), must be ABO compatible with patient; usually compatibility tests are performed prior to issue.

➢ Transfuse as soon as possible after collection.

➢ Use standard blood filter (170 µ screen) set, do not use LR or microaggregate filters (20 µ screen).

❑ Quality Control Requirements: ≥ 1.0x1010 granulocytes in at least 75% of units tested

Granulocyte Apheresis - Characteristics

❑ Volume:

❑ Storage Temp:

❑ Expiration:

❑ Transport Temp:

❑ Contents:

❑ Limitations:

❑ Quality Control Requirements:

- Crystalloids

- Colloids

What are the common intravenous (IV) solutions?

❑ Factor VIII & vWF (Alphanate®) - treat Hemophilia A and von Willebrand Factors diseases

❑ Factor IX (Konyne®) - treat Hemophilia B

❑ Gamma (IgG) globulin (Gammagard®) - treat Ig-associated disease

What are some examples of the blood-derivative pharmaceutical products?