Rejuvenation 4.
Development of RBC substitutes
It is the process by which ATP & 2,3-DPG levels are
restored or enhanced by metabolic alterations.
Add info: 2,3 -Diphospoglycerate is important in
oxygen binding curve
◼ Uses rejuvenation solutions such as:
1. PIGPA = phosphate, inosine, glucose, pyruvate &
adenine.
2. PIPA
3. Rejuvesol = the only FDA-approved rejuvenation
solution in the US.
How is rejuvenation done?
The RBC unit is incubated at 300C for 1H with 50mL
rejuvenation solution.
◼ RBCs are then washed to remove nonmetabolized
rejuvenation solution materials & deleterious
amounts of extracellular K+.
◼ Rejuvenated RBCs are used within 24H of thawing
to prevent the entrance of bacteria.
◼ Expensive & time-consuming, therefore, it is not
used often but is invaluable for preserving selected
autologous & rare units of blood for later use.
Add info: expensive and time consuming ang
rejuvenation process
Current Trends in Blood/RBC Preservation
Research:
Research & development in RBC preparation &
preservation is being pursued in 4 directions:
1. Development of modified/new additive solutions
2. Development of procedures to reduce the level of
pathogens that may be in RBC units
3. Development of procedures to convert A(-), B(-),
& AB(-) RBCs to O(-) RBCs
Add info:
*not all rbcs are available mas mayo if nay rbcs
substitute.
*if A(-), B(-), & AB(-) RBCs are not available convert
to O nalang, oten
Platelet Preservation
Role of platelets in hemostasis:
1. Initial arrest of bleeding by platelet plug
formation = involves adhesion of platelets to the
endothelium & subsequent aggregation with
thrombin.
2. Stabilization of the hemostatic plug by
contributing to the process of fibrin formation &
maintenance of vascular integrity.
Add info:
Hemostasis – stoppage or stopping of a flow of
blood
Homeostasis – maintain balance in our body
2 stages: primary hemostasis – your platelets are
the one who are active, although the platelet plug
formation is not stable but still maka stop gihapon
sa flow sa blood.
Kugang- platelet plug
Secondary hemostasis – clotting factor and fibrin
formation
Clinical Use of Platelets:
1. To treat bleeding associated with
thrombocytopenia
2. To prevent bleeding secondary to drug &
radiation therapy
Methods of preparing platelets for transfusion:
1. Platelet-rich-plasma (PRP)-derived
 2. Whole blood-derived platelet concentrates (PC)
3. Apheresis
PRP-derived platelets:
◼ Approximately 250-300mL in volume.
◼ Circulatory overload is the major complication.
Whole blood-derived PC:
➢ Prepared from units of whole blood drawn into
triple or quad collection bag systems.
➢ Harvesting of platelets from whole blood is
done using
2 methods:
1. PRP method = used in North America
2. Buffy coat method = used in Europe
How to obtain PRP-derived platelets?
◼ Units of PRP are separated by a slow
centrifugation step.
◼ PRP is centrifuged again with hard spin
conditions to concentrate the platelets.
How to obtain platelets in the buffy coat
method?
➢ Units of whole blood are centrifuged with hard
spin conditions to concentrate the platelets in the
buffy coats.
➢ Single or pooled buffy coats are centrifuged with
slow spin conditions harvesting the platelets in the
plasma supernatant.
PRP-derived platelets:
◼ Approximately 50mL plasma (range: 45-65mL) is
retained with the platelets in order to maintain a pH
of at least 6.2 during storage.
◼ Storage is 5 days at 200C to 240C (RT).
◼ Expiration is midnight of day 5.
◼ 1 unit may contain 5.5 x 1010 platelets.
◼ if the seal of any PC bag is broken, the platelets
need to be transfused within 4H when stored at RT.
Plateletpheresis:
◼ Platelets are harvested by drawing blood from a
donor into an apheresis instrument (cell separator)
◼ The instrument separates the blood into
components using centrifugation.
◼ Platelets are retained.
◼ Remainder of the blood is returned to the donor.
◼ Platelets are then concentrated in storage
containers after repeated cycles.
◼ Apheresis components contain at least 4-6 x as
many platelets obtained from whole blood (3-4 x
1011 platelets)
Platelet storage:
◼ PC prepared from WB & apheresis are routinely
stored at RT with constant agitation for up to 5
days.
◼ Flat-bed or circular agitators are available.
◼ Although they can be stored at 1-60C, it is not
routinely done.
Reasons why platelets are not preserved in
cold storage:
1. Cold temperature converts the normal discoid
shape of platelets to irreversibly spherical.
2. Microtubule disassembly.

Why is it necessary to agitate PC?

1. To facilitate O2 transfer into the platelet bag.
2. To facilitate O2 consumption by the platelets.
3. To prevent aggregation or clumping.
Platelet Storage & Bacterial Contamination:
➢ Bacterial growth is the major concern with
platelet storage at RT.
➢ Some reasons for bacterial contamination
include:
1. Unremoved contamination at the phlebotomy
site.
2. The donor has unrecognized bacterial infection.

Current Trends in Platelet Preservation

Research:
1. New documentation of platelet storage for 7
days with testing for bacteria.
2. Development of additive solutions or synthetic
media.
3. Development of procedures to reduce the level
of pathogens in the platelet units.
4. Development of platelet substitutes.
5. New approaches for platelet storage at 1-60C.
6. Development of cryopreserved platelets.
Add info: sa manila ang mhar gen kay 5 days
gihapon ang storage

Basa rang uban….end