RBC MEMBRANE

• Semi-permiable lipid bilayer supported by a

meshlike protein cytoskeleton structure.

• Main lipid components of the membrane are

phospholipids.

• Consists of 2 kinds of protein : Integral Proteins &

Peripheral Proteins.
 RBC
CHARACTERISTICS
Deformability
RBC flexibility and deformability
 Exemplified by the formation of spherocytosis & bite
cells

Permeability
• Permeable to water & anions (Chloride & bicarbonate)
• Impermeable to cations (Sodium & Potassium)
 METABOLIC PATHWAYS OF
RBC
1. Anaerobic Glycolytic Pathway
 Generates about 90% of ATP needed by the RBC

2. Pentose Phosphate Pathway

 Produce approx. 10% of the ATP needed by the RBC
 Production of glutathione

3. Methemoglobin Reductase Pathway

 Maintain iron in the ferrous functional state
 A defect can affect RBC post-transfusion survival and function

4. Luebering Rapoport Shunt

 Permits the accumulation of 2,3-DPG
RBC PRESERVATION

SUCCESFUL TRANSFUSION:
 75% of post-transfusion RBCs remain viable for 24hours.

2 CRITERIA TO EVALUATE NEW PRESERVATION

SOLUTIONS AND STORAGE CONTAINERS (FDA):
 An average 24-hour post-transfusion RBC survival of more
than 75%
 Free hemoglobin <1% of total hemoglobin
 RBC STORAGE
LESION
Characteristic Change Observed
% Viable cells

Glucose

ATP DECREASED
pH

2,3-DPG

Oxygen Dissociation Curve Shift to the LEFT

Plasma K+

Plasma Hemoglobin INCREASED

Lactic Acid
 Plasma
hemoglobin
Plasma K+
Na+ Lactic Acid
K+

Viable cells
Glucose
pH
Helps release oxygen
ATP from hemoglobin (once
transfused, ATP & 2,3-
2,3-DPG DPG return to normal)
Plasma Na+
 RBC PRESERVATION
1. ANTICOAGULANT PRESERVATIVE SOLUTIONS

APPROVED ANTICOAGULANT PRESERVATIVE SOLUTIONS

NAME ABBREVIATION STORAGE TIME

(DAYS)
Acid citrate-dextrose (formula A)* ACD-A 21
Citrate-phosphate-dextrose CPD 21
Citrate-phospate-double-dextrose CP2D 21
Citrate-phosphate-dextrose- CPDA-1 35
adenine
*ACD-A is used for apheresis components
 CHEMICALS IN ANTICOAGULANT SOLUTIONS

CHEMICAL FUNCTION PRESENT IN

ACD-A CPD CP2D CPDA

Citrate (Sodium Chelates calcium ; Prevents X x X X

Citrate/Citric Acid) clotting
Monobasic Maintains pH during X X X X
sodium storage; Necessary for
phosphate maintenance of adequate
levels of 2,3-DPG
Dextrose Substrate for ATP x X x X
production (cellular energy)
Adenine Production of ATP (extends x
shelf-life from 21-35 days)
 RBC PRESERVATION
2. ADDITIVE SOLUTIONS
 Added to RBCs after removal of the plasma with or
without platelets.
 AS is contained in a satellite bag.
 Benefits of additive solutions:
 Extends the shelf-life
 Harvesting of more plasma & platelets
 RBC concentrate of lower viscosity
 ADDITIVE SOLUTIONS LICENSED IN THE US

NAME ABBREVIATION STORAGE TIME (DAYS)

Adsol (Baxter Healthcare) AS-1 42
Nutricel (Pall Corporation) AS-3 42

Optisol (Terumo AS-5 42

Corporation)

In general, the AS employed are composed of :

• Saline (S)
• Adenine (A)
• Dextrose or Glucose (G)
 AS-1 & AS-5 contains mannitol (to retard hemolysis).
 AS-3 contains citrate & phosphate for the same purpose.
 RBC PRESERVATION
3. RBC FREEZING
 Primarily used for autologous units and the storage of rare
blood types.
 Involves the addition of cryoprotective agent to RBCs that
are less than 6 days old.
 The FDA licenses frozen RBCs for a period of 10 years
from the date of freezing.
 Glycerol is the most commonly used.
 2 concentrations of glycerol:
1. High-concentration glycerol (40% w/v)
2. Low-concentration glycerol (20% w/v)
ADVANTAGES OF HIGH-CONC. GLYCEROL OVER LOW-CONC. GLYCEROL

ADVANTAGE HIGH GYCEROL LOW GLYCEROL

Initial freezing temperature -80OC -196 OC

Need to control freezing rate No Yes

Type of freezer Mechanical Liquid Nitrogen

Maximum storage -65 OC -120 OC

temperature
Shipping requirements Dry Ice Liquid Nitrogen

Effect of changes in storage Can be thawed and Critical

temperature refrozen
RBC PRESERVATION
4. RBC REJUVENATION
o RBCs in the liquid state can be rejuvenated at
outdate or up to 3 days after outdate by
incubation for 1-4 hours at 37oC with
rejuvenation solution.
o Rejuvesol (enCyte Systems) is the only FDA-
approved rejuvenation solution. It consists of
phosphate, inosine, pyruvate, and adenine
(PIPA).
RBC PRESERVATION
5. RBC SUBSTITUTES
2 categories:
1.Hemoglobin-based oxygen carriers
– include stroma free hemoglobin solution (SFHS),
chemically modified hemoglobin solution, recombinant
hemoglobin, and encapsulated hemoglobin.

2. Perfluorochemicals
– chemically inert but excellent gas solvents.
– carry O2 and CO2 by dissolving as much as 40% to
70% oxygen per unit volume.
 Red Cell Antigen-
Antibody Interactions
1. Agglutination
 Clumping of red cells due to antibody coating
2. Hemolysis
 Direct lysis of RBCs due to antibody coating
and subsequent complement fixation
3. Precipitation
 Formation of insoluble complex when soluble
antigen reacts with soluble antibody
 Grading of Agglutination
Reactions
+4 One Solid aggregates, clear background

+3 Several large aggregates, clear background

+2 Many medium size aggregates, clear background

+1 A few small aggregates, cloudy background

+/- Tiny aggregates barely visible

0 No agglutination

mf Mixed Field

H Hemolysis

PH Partial Hemolysis
 Basic Genetics
• Genetics - the study of inheritance or
the transmission of characteristics
from parents to offspring.
• Gene – basic unit of genetic
information.
• Chromosomes – storage units of genes.
• Locus – location of a gene/marker on
the chromosome.
• Allele – one variant form of a
gene/marker at a particular locus.
 Sex Chromosomes vs.
Autosomes
Most human cells
contain 46 chromosomes:

• 2 sex chromosomes (X,Y):

XY – in males.
XX – in females.

• 22 pairs of chromosomes named autosomes.

 Phenoype vs. Genotype

• Genotype - the genetic makeup of an

organisM.
• Phenotype - the visible properties of an
organism.
Dominant vs. Recessive
Genes
• “A” allele “B” allele is dominant.

• Together though, the “A” and “B”

alleles are co-dominant.

• “O” allele is recessive.

Determining the Genotype

Punnett Squares
Mother’s Genotype

Father’s
Genotype
 Scenario 1

Suppose that a mother has blood Type A

and genotype AO and the father has blood
Type B and genotype BO.

Draw a Punnett square to show the possible

genotypes of their children.

What are the phenotypes of the kids?

 Review

• What are perfluorocarbons (PFC)?

a) Rejuvenation solution
b) Additive
c) Anticoagulant
d) Blood Substitute
 Review

• Storage time of CPDA-1

a) 21 days
b) 35 days
c) 42 days
d) 10 years
 Review

• Storage time of Adsol

a) 21 days
b) 35 days
c) 42 days
d) 10 years
 Review

• Generates about 90% of ATP needed by the

RBC
a)Anaerobic Glycolytic Pathway
b)Pentose Phosphate Pathway
c) Methemoglobin Reductase Pathway
d)Luebering Rapoport Shunt