1 RBC AND PLATELET PRESERVATION: Historical Perspective and Trends

I. INTRODUCTION

IMMUNOHEMATOLOGY: immunologic properties and reaction of all blood components

BLOOD BANK: organized unit responsible for collecting processing and storing blood for transfusion purposes; hospital or non-hospital based (A,B,C,D)
BLOOD: complex liquid comprising various cellular components suspended in aqueous solution; trbsport

II. HISTORY

1492 Pope Innocent VII- first recorded transfusion in history World War II Charles Drew
EdwardLindemann Vein- to- vein transfusion with multiple syringr and cannula for Dev technique in blood transfusion and preservation (spread of
puncturing vein bb)
1869 Braxton Hicks{ sodium phosphate Stimulated blood preservation research
1901 Karl Landsteiner- discovered ABO blood group 1941 Dr. Charles Drew: appointed director of the first American red
1914 Hustin- sodium citrate (less acidic) Cross blood bank at Presbyterian Hospital
1915 Lewisohn- minimum amt of citrate needed for anticoagulation 1943 Loutit and Mollison: formula for preservative acid-citrate
450 ml = 63ml dextrose (fopr calcium ions, di mag accumulate)
500-550 ml= 70ml July 1947 Journal of Clinical Investigation
250 ml – 43-44 ml 1957 Gibson: : citrate phosphate dextrose
1916 Rous- Turner: citrate dextrose solution for RBC preservation Citrate= chelate Ca
(prolong shelf life) Phosphorus= generate ATP for RBC

III. COMPONENTS OF BLOOD

 Apheresis: collecting platelets from whole blood

 Cryoprecipitation: converting plasma into clotting factor concentrate rich in antihemophilic fact

Whole blood collection

- 450ml (63 ml preservative)
- 500 ml (70 ml; Modified plastic collection)
- 525 ml (maximum volume collected from 110 pound donor)
- Donor can replenish lost fluids after 24 hours
- Donated rbcs replaced within 1-2 months after donation
- Volunteer donor can donate whole blood every 8 weeks

3 STEPS IN BLOOD DONATION

1. Educational Reading Material Transfusion Transmitted Infection –

TTI)
2. Donor Health History Questionnaire
3. Physical Exam

IV. RBC BOLOGY AND PRESERVATION

RBC CHEMICAL COMPOSITION PHYSICAL COMPOSITION

1. Iron, Hgb 1. Unnucleated
2. Protein 2. Biconcave
3. Lipid 3. 7um
4. FFA, CF

Normal Erythrocyte Function:

1. Normal chemical composition and structure
2. Hemoglobin structure and function
3. RBC metabolism
Decreased Deformabilty: Permeability
1. Low ATP Prevent colloid hemolysis and control the volume of RBC
 Decreased phosporylation of 1. Permeable to water and anions
spectrin; 2. Impermeable to cations
2. High Calcium  Active transport
 Increased membrane rigidity  Intracellular and extracellular ratio of Na and K , respectively 1:12, 25:1
3. Loss of RBC membrane (spherocyte,  Calmodulin- cytoplasmic calcium-binding protein, control s the pumps and prevents excessive Ca in cell;
bite cell) When ATP depleted, Ca and Na accumulate within cell, K and water are lost, resulting in dehydrated rigid cell
 2 RBC AND PLATELET PRESERVATION: Historical Perspective and Trends

METABOLISM RBC metabolism divided into 4 (1 glycolytic pw and 3 ancillary pathway) Process for Normal hemoglobin dependent on :
1. Embden-Meyerhof Pathway (Glycolysis) 1. Adequate iron delivery and supply
 Generates 90% of the ATP needed by the RBC 2. Adequate synthesis of protoporphyrin (presence of heme)
3. Adequate globin synthesis
2. Phosphogluconate Pathway (pentose-phosphate pw)
Deoxyhemoglobin/ Tense form
 Provides 10% of ATP needed Unloading; low affinity of O2 in tissue
3. Methemoglobin reductase pathway 1. Widen space between B chain
 Maintain heme iron of hemoglobin in ferrous fnc state 2. Bind 2,3 –DPG
 Defect can affect RBC post transfusion survival and function 3. Salt bridges
Oxyhemoglobin/ Relaxed form
4. Leubering-Rapaport Pathway Loading; high O2 affinity in lungs
 Accumulation of 2,3-DPG necessary for the affinity of HgB for O2 1. Saltbridges broken
2. B chain pulled together
3. Expel 2,3- DPG

Hemoglobin-Oxygen Dissociation Curve

1. Hgb primary function: gas transport; O2 to tissue, CO2 to lungs/excreted
2. Sigmoid-curve relationship
3. Shift to the right- decrease affinity to o2 and increase delivery to tissue; hypoxia
4. Shift to the left- increased affinity to o2 and decreased delivery to tissue
PRESERVATION  The goal of preservation is to provide viable and functional blood components for patients requiring blood transfusions.
 Criteria used to evaluate new preservation solutions and storage containers:
1. Average 24-hour post-transfusion RBC survival of >75%
2. Cell rigidity be maintained throughout the shelf-life of the stored RBC
 To maintain optimum viability, blood must be stored in liquid state between 1 to 6 degrees for a specific number of days.
 LESION OF STORAGE: loss of rbc viability, can be restored via salvaging.

Anticoagulant preservation Solution

 Used as an attempt to stimulate glycolysis so that ATP levels were better maintained.
 Blood stored in CPD preservatives also becomes depleted of 2,3-DPG by the second week of storage
 Pathophysiological effects of transfusion of rbcs with low 2,3-DPG leels and increased affinity for oxygen include:
- Increase in cardiac output
- Decrease in mixed venous (pO2) tension
 Stored rbcs regain the ability to synthesize 2,3-DPG after transfusion; approximately after 24 hours normal levels are restored

Additive Solutions
 Preserving solutions added to the RBCs after removal of the plasma with or without platelets.
 This was made due to the loss of nutrients needed to maintain RBC during
storage after the removal of plasma components
 Benefits include:
- Extending shelf-life of RBC
- Allows harvesting more plasma and platelets from the unit
- Produces an RBC concentrate of lower viscosity that is easier to
infuse
 100 ml of additive solution to 450 ml blood collection
 Final hematocrit from 70-80% to around 50-60%
 None of the additive solutions maintain 2,3-PDG throughout the storage time. As with RBCs are stored only with primary anticoagulant preservatives, 2,3-DPG is
depleted by the second week of storage
 3 RBC AND PLATELET PRESERVATION: Historical Perspective and Trends

RBC Freezing
 Used for autologous units and storage of rare blood types (autologous transfusion:
donating for self use)
 Glycerol is added to RBCs that are less than 6 days old slowly with vigorous shaking
 High concentration glycerol, 40%; low concentration glycerol, 20%

Deglycerolyzation process
- Achieved by systematically replacing the cryoprotectant with decreasing
concentrations of saline: 12% - 1.6 %- 0.2% dextrose in normal saline.
- Outdating period:
a. Open system: 24 hours at 1-6 degrees
b. Closed system: 15 days at 1-6 degrees ( ACP 215, Haemonetics)

RBC Rejuvination
 Process by which ATP and 2,3-DPG levels are restored or enhanced by metabolic
alterations
 Rejuvesol: phosphate, adenine, inosine, pyruvate
 Approved for use with CPD, CPDA-1, CPD/-AS1
 RBCs stored in liquid state can be rejuvenated at outdate or up to 3 days after outdate,
depending on rbc preservative solution used
 Only 450ml collections can be rejuvenated
 Steps:
- Rejuvenate rbc with 50ml rejuvenating solution at 37 deg for 1 hour
- Rbcs are then washed and can be transfused

GENETICS
1. Genetics- inheritance/ transmission of characteristics from parents to offspring
2. Genes- hereditary units
3. Chromosomes- strings of beads on a strand of DNA
4. Locus- location of chromosome in genes
5. Allele- several expressions/ forms of the gene
6. Phenotype- outward expression/ observable characteristics
7. Genotype- notation of the actual genes inherited; complete genetic constitution of a group/ organization
8. Homozygous- Inherited identical alleles
9. Heterozygous- inherited different alleles
10. Amorph- silent gene; does not produice any detectable traits
11. Dominant trait- expressed trait
12. Recessive Trait- expressed if homozygous; normally non expressed gene
13. Humans- 46 chromosomes (23 pairs); 22 autosome, 1 set sex chromosome
14. ABO BGS- gene that codes is located at long arm of chromosome 9 terminal portion