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BLOOD COMPONENT PREPARATION,
STORAGE AND USE
BY
Salvatory Temu
18/12/2023
BLOOD COMPONENTS
• Blood components, including packed red blood
cell (PRBC) concentrate, fresh frozen plasma and
cryoprecipitate, platelet concentrate and
granulocyte products.
• Prepared either from whole blood donation or by
automated apheresis.
• Blood component preparation and manufacture
allow each component to be manufactured and
stored under optimal conditions.
Apheresis
BLOOD COMPONENTS
• Apheresis is a procedure where required single or
more than one component is collected, and the
rest of blood components are returned back to
the donor.
• The working principle of apheresis equipment is
either by centrifugation (different specific gravity)
or by filtration (different size).
BLOOD COMPONENTS
• The use of components (i.e. component
therapy) has largely replaced the use of whole
blood due to the ability to choose individual
components and component constituents that
target a specific patient’s needs as well as
enabling each component to be stored
optimally.
BLOOD COMPONENTS
• The blood components being foreign to a patient may
produce adverse effects that may range from mild
allergic manifestations to fatal reactions.
• Such reactions are usually caused by plasma proteins,
leucocytes, red cell antigens, plasma and other
pathogens.
• To avoid and reduce such complications, blood
products are modified as leukoreduced products,
irradiated products, volume reduced products, saline
washed products and pathogen inactivated products.
BLOOD COMPONENTS
Primary blood bag with satellite bags
BLOOD COMPONENTS
Key terminologies in blood preparation
• Whole blood: blood collected before separation into
components.
• Components: parts of whole blood that are separated.
• Closed system: a sterile system of blood collection.
• Open system: when the collection is exposed to air,
decreasing expiration date.
Collection of Blood and blood components
• Blood is collected into a primary bag containing anticoagulant-
preservatives and depending on the components choice;
satellite bags with hermetic seal in between may also be
attached.
• The blood bag anticoagulant-preservative minimizes
biochemical changes and increases shelf life of the component
or whole blood.
• Blood unit is collected into a closed system within 15 minutes to
avoid activating the coagulation system.
• Transfuse within 24 hours if the hermetic seal is broken and
stored at 1-4˚C, or within 4 hours if stored at 20-24˚C.
Whole Blood (WB)
• Clinical indications for use of WB are now becoming extremely limited. Whole blood
comprises RBCs, WBCs, platelets and plasma (with anticoagulant).
• Used for massive transfusion to correct acute hypovolaemia such as trauma and shock,
exchange transfusion.
• 1 unit increases Hgb 1 g/dL and Hct 3%
• RARELY used today, platelets non-functional, labile coagulation factors gone.
• Must be ABO identical.
Whole Blood
• Whole blood is the most common starting
product for component preparation and
manufacture.
• Whole blood is generally not found in most blood
banks today because component therapy is more
appropriate to target a patient’s specific
indications for transfusion (for example, RBC
products for symptomatic anemia, plasma
products for multiple coagulation factor
deficiencies, and platelet products for
thrombocytopenia or platelet dysfunction).
• In addition, in whole blood, platelet function
and post-transfusion survivability are lost
when the whole blood is stored in the
refrigerator, and coagulation factor activity,
especially for labile factors (factors V and VIII),
deteriorates over time.
• In general, 500 ml of whole blood is collected
into a bag with 70 ml of anticoagulant-
preservative solution (see below), creating a
product with a final hematocrit of ~38%.
• Whole blood is stored at 1–6°C.
• Depending on the manufacturer’s system and
a country’s regulations, different hold times
are allowed before the preparation of blood
components from whole blood.
• If platelet products are to be manufactured
from whole blood, the whole blood product
must be stored at room temperature until the
platelets are removed.
• Fresh whole blood is occasionally used by the
US military via ‘walking blood donors’ (donors
who are ABO/D typed, infectious disease
tested and available to donate in times of
need).
• This product is transfused as soon as possible
and has a shelf-life of 24 hours when stored at
room temperature, allowing maintenance of
platelet function, while still minimizing the risk
of bacterial overgrowth.
• The primary use of this product is to infuse
viable platelets when other platelet products
are not available.
• Risks associated with use of this product
include transfusion-transmitted diseases and
transfusion associated graft versus host
disease.
• Blood component preparation was developed
in 1960 to separate blood products from one
unit whole blood by a specialised equipment
called as refrigerated centrifuge.
Component Manufacturing
• When whole blood is manufactured into
components, it is collected into a primary bag
containing an anticoagulant-preservative
solution.
• The primary bag has up to three attached
satellite bags for RBC, platelet, and plasma or
cryoprecipitate component manufacturing.
Diagrammatic representation of blood components preparation
• Due to the different specific gravities of RBCs
(1.08–1.09), plasma (1.03–1.04) and platelets
(1.023), differential centrifugation of the
whole blood unit is used to prepare blood
components.
• Optimal component separation requires
specific centrifugation variables, such as rotor
size, speed, and duration of spin (Figure 9.1).
• Preparing only PRBC and fresh frozen
plasma(FFP) is by single-step heavy spin
centrifugation; however preparing platelet
concentrates(PLTCs), PRBC concentrates and
FFP is by two step centrifugation.
Anticoagulant-Preservative Solutions
• Whole blood is collected into containers with
anticoagulant-preservative solutions, including
acid-citrate-dextrose (ACD), citrate-phosphate-
dextrose (CPD), citrate-phosphate-dextrose-
dextrose (CP2D) and citrate-phosphate-dextrose-
adenine (CPDA-1).
• The citrate (sodium citrate and citric acid) acts as
an anticoagulant, and the phosphate (monobasic
sodium phosphate and trisodium phosphate),
adenine and dextrose are substrates for cellular
metabolism.
Packed RBC Components
• RBC products are used primarily for the treatment of
symptomatic anemia or hemorrhage in order to
increase tissue oxygenation.
• RBC products are prepared either by centrifugation of
whole blood followed by removal of 200- 250ml of
platelet-rich plasma layer, or by automated apheresis
collection.
• Anticoagulant preservative solutions allow RBC
components to be stored for extended periods of time
at 1–6°C without a significant detrimental effect on the
quality of the RBC.
• Prepared by sedimenting whole blood (WB) or centrifuging it to
remove 200-250ml of supernatant plasma from a unit of WB.
• This produces 200-250ml red cell concentrates with a haematocrit
of approximately 80% for non-additive (CPD), 60% for additive
(ADSOL).
• A unit increases patient’s Hb level by about 1g/dL (10g/L) and
haematocrit by 3%.
• It does not contain functional platelets or granulocytes and has
same O2 carrying capacity with W.B.
• It is used to treat symptomatic anaemia and routine blood loss
during surgery to increase patient’s red cell mass without
increasing their blood volume.
• The shelf-life of a given product is determined
by the solution used and based on the
criterion that there is less than 1% hemolysis
in the product at the end of storage, and 75%
of the transfused RBCs remain viable 24 hours
after transfusion.
• RBCs stored in CPD or CP2D have a shelf-life of
21 days and in CPDA-1 of 35 days.
• RBCs stored in CPD, CP2D or CPDA-1 have a
hematocrit of ~80% with a final volume of
225–350 ml and up to 110 ml of plasma.
• The majority of RBC products are stored in
additive solutions, which extend the shelf-life
of the product to 42 days.
Packed red cell concentrate: Once the unit of red cell
concentrate is “opened”, it has a 24 hour expiration date.
Additive Solutions
• Additive solutions, such as AS-1, AS-3 and AS-
5, can be added to the primary anticoagulant-
preservative solution to increase the shelf-life
of RBCs to 42 days.
• All of these solutions contain dextrose,
adenine and sodium chloride; AS-1 and AS-5
contain mannitol, and AS-3 contains
monobasic sodium phosphate, sodium citrate
and citric acid.
• Additive solution products have a final volume of 350–
400 ml and hematocrit of 55–65%, containing 100–110
ml of additive solution and 10–40 ml of plasma.
• In general, additive solutions must be added to the
product within 24 hours of collection, depending on
manufacturer’s specifications.
• Outside of the US, SAGM (saline-adenine-glucose-
mannitol), which is hypertonic, is used and PAGGSM
(phosphate-adenine-glucose-guanosin-saline-
mannitol), which is isotonic, is under development.
• A number of other additive solutions are
currently under study, including EAS-81
(NaHCO3, Na2HPO4, adenine, dextrose, and
mannitol) as well as methods to decrease
oxidative damage by adding GSH precursors
(glutamine, glycine and N-acetyl-L-cysteine) or
O2 removal.
RBC Modification
• RBC products can be further modified for
specific patients’ needs.
• Multiple modifications can be performed on a
single product.
• Examples of product modifications and their
use include the following:
• Freezing: Frozen RBC products, which are
cryopreserved with glycerol, have an extended storage
life of up to 10 years when stored at −65°C or below.
• The primary indications for frozen RBCs are for rare
RBC phenotypes (e.g. products from donors who lack
high frequency RBC antigens) or autologous donations,
or to maintain product inventory for disaster
management.
• Frozen RBC products must be deglycerolized prior to
transfusion.
• Deglycerolized products have a 24-hour outdate.
• Volume reduction: RBC products can be volume-
reduced to remove most of the supernatant.
• The primary indications are to remove the
accumulated potassium, which leaks from the
RBCs during storage, in order to prevent
hyperkalemia in at-risk patients such as neonates;
to reduce the AS concentration, though this is
rarely required or appropriate; or to reduce the
volume infused in volume-sensitive patients.
• Washing: RBC products can be washed to
remove the plasma and, with it, plasma
proteins.
• This process can be used to minimize the risk
of recurrence of severe allergic/anaphylactic
reactions.
• Rejuvenation: Rejuvenated RBCs have 2,3-DPG
and ATP levels near to those of a freshly drawn
unit.
• During storage 2,3-DPG levels fall linearly
towards zero during the first two weeks.
• ATP levels fall to 50–70% of initial levels during
storage, depending on the anticoagulant-
preservative solution used.
• Rejuvenation is usually performed prior to
freezing RBCs.
• The RBCs must be washed prior to use to
remove the rejuvenation solution.
• Leukoreduction: Leukoreduced RBCs are used
to mitigate the risk of febrile nonhemolytic
transfusion reactions, HLA alloimmunization,
CMV transmission and transfusion-related
immunomodulation, each in appropriate and
clinically indicated populations.
• Most institutions and clinicians prefer
universally available leukoreduced products.
• Irradiation: Irradiated RBCs are used to
prevent transfusion-associated graft versus
host disease.
• Aliquots: RBC products can be divided into
smaller volumes for neonatal transfusions.
Plasma Components
• Plasma components are primarily used to treat
multiple plasma factor deficiencies or to prevent
development of a coagulopathy.
• Plasma contains all of the coagulation factors found in
whole blood and the critical proteins albumin and
fibrinogen.
• Plasma products may be manufactured either from
whole blood or by automated apheresis.
• Plasma products that are used in the transfusion
service include fresh frozen plasma (FFP), plasma
frozen within 24 hours of phlebotomy (FP24) and
thawed plasma (TP).
• FFP must be frozen within 6–8 hours of collection,
while FP24 is frozen between 8 and 24 hours of
collection.
• FP24 has lower levels of factors V and VIII than FFP, but
this reduction is rarely of significance.
• FFP and FP24 are stored at −18°C or colder for up to
one year (with approval from the FDA, FFP can be
stored at −65°C or colder for up to seven years).
• Prior to transfusion, plasma must be thawed at 30–
37°C, which requires ~20 minutes; it is then stored at
1–6°C if not transfused immediately.
• Once thawed, FFP and FP24 must be
transfused within 24 hours, otherwise it
becomes classified as TP, which may be stored
at 1–6°C for up to five days.
• During these five days of storage, factor VIII
activity levels decrease by approximately 30%.
• FFP, FP24 and TP can be used interchangeably
in most situations.
• Plasma products may be further
manufactured into cryoprecipitate and
cryoprecipitate-reduced plasma.
• In addition, outside of the US, large pools of
plasma are manufactured into
solvent/detergent-treated plasma, which has
minimal (if any) risk of transmitting lipid-
enveloped viruses (HIV, HCV, and HBV).
• Other pathogen inactivated plasma products
available outside of the US, which are not
pooled, include methylene blue (MB),
riboflavin and ultraviolet light, and
amotosalen and ultraviolet light though some
manufacturers and countries have considered
removing MB from use due to allergic
reactions.
• Plasma (stored at −18°C or below) and liquid
plasma (stored at 1–6°C) are manufactured from
whole blood no later than five days after the
whole blood expiration date, and are converted
into an unlicensed product termed ‘recovered
plasma’ that can be shipped for further
fractionation into albumin, antithrombin III,
factor VIII or factor IX concentrates, or
immunoglobulin preparations.
• Liquid plasma is sometimes, though rarely, used
for transfusion.
Fresh Frozen Plasma
• Prepared by removing plasma from WB within 8 hours of
collection and must be frozen within same duration.
• It is used to replace labile and non-labile coagulation factors
in massively bleeding patients and treat bleeding associated
with clotting factor deficiencies when factor concentrate is
not available.
• Its constituents are water, carbohydrates, fats, minerals,
proteins (all labile and stable clotting factors).
• Each unit of FFP measures 200-225ml and elevates the level
of each clotting factor by 2-3% in adults, the therapeutic
dose is 10-15ml/kg.
Fresh Frozen Plasma
• The storage temperatures:
- frozen -18˚C, preferably -30˚C or lower
- thawed - 1-6˚C
- Thawed in 30-37˚C water bath.
• Expiration:
- frozen - 1 year if stored at <-18˚C.
- frozen - 7 years if stored at <-65˚C.
- thawed - 24 hours.
• Must have mechanism to detect units which have thawed and
refrozen due to improper storage.
• Must be ABO compatible.
• FFP is thawed before transfusion at 30-37°C in a waterbath
for 30-45 minutes and can then be stored at 1-6°C and
transfused within 24 hours.
Fresh Frozen Plasma
Fresh Frozen Plasma
• It is indicated for use in the following conditions:
- Replacement for isolated coagulation function deficiencies.
- The reversal of warfarin therapies.
- In the case of massive blood transfusion.
- Antithrombin III deficiency treatment.
- Correction of coagulopathy or liver disease.
- Thrombotic thrombocytopenic purpura.
Cryoprecipitated antihemophilic factor
(AHF)
• The primary indication for the use of
cryoprecipitate (the FDA term is cryoprecipitated
AHF) is for replacement of fibrinogen.
• Cryoprecipitate is prepared by slowly thawing FFP
to 1–6°C, thereby leading to formation of a
precipitate, which is collected and refrozen to
make cryoprecipitate.
• Each unit of cryoprecipitate, which is
approximately 15 ml, must contain fibrinogen
(>150 mg) and factor VIII (>80 IU) at these
specified levels.
• In addition, each unit contains von Willebrand’s factor (vWF) (80–
120 IU), factor XIII (40–60 IU) and fibronectin.
• Cryoprecipitate is usually administered to adults in doses of 8–10
donor units, which are pooled prior to transfusion (known as a
‘cryopool’).
• Some blood centers manufacture prepooled cryoprecipitate, where
five units are pooled together prior to storage.
• This product is easier for the transfusion service to use in cases of
emergency than pooling prior to issue (post-thaw).
• Cryoprecipitate units can be stored for up to one year at −18°C and
must be transfused within four hours of pooling using an open
system and within six hours of thawing using a closed system.
Cryoprecipitated antihemophilic factor (AHF)
• Cryoprecipitated antihemophilic factor (AHF) or “Cryo” is the
cold-insoluble portion of plasma that precipitates when FFP
is thawed at 1-6˚C.
• It contains high levels of Factor VIII (about 80 IU in each
unit), plasma (10-15ml), von Willebrand’s factor (platelet
adhesion), Fibrinonectin and Fibrinogen (150 mg in each
unit).
• It is used for treatment of hemophiliacs and Von Willebrand
disease when concentrates are unavailable.
• Used most commonly for patients with DIC or low fibrinogen
levels and the therapeutic dose for an adult is 6 to 10 units.
Cryoprecipitated antihemophilic factor (AHF)
• Can be prepared from WB which is then designated as
"Whole Blood Cryoprecipitate Removed" or from FFP.
• The plasma is first frozen, then thawed at 1-6˚C which results
in the formation of a precipitate.
• The plasma is centrifuged, cryoprecipitate goes to the
bottom.
• Plasma is then expelled or removed and frozen at -18 ˚C
within 1 hour of preparation.
FFP
Frozen
within 8
hours
Thawed
FFP
Cryoprecipitate
(VIII, vW)
Plasma cryoprecipitate reduced
(TTP, FII, V, Vii, IX, X, XI)
Thaw at 30-37°C
Store at RT 4 hrs
Refrozen with 24 hrs of
separation
Store at ≤18°C 1 yr
5 day expiration at 1-6°C
Cryoprecipitated antihemophilic factor (AHF)
• Same storage as FFP (cannot be re-frozen as FFP once it is
separated); at -18˚C for 1 year and if thawed, store at room
temperature for 4 hours.
• The leftover plasma is called “cryoprecipitate-reduced” or
"plasma cryo".
• Cryoprecipitate-reduced used for thrombotic
thrombocytopenic purpura (TTP), 2° treatment for Factor VIII
deficiency (Hemophilia A), 2 ° treatment for von Willebrand’s
Disease, Congenital or acquired fibrinogen deficiency, FXIII
deficiency and “Fibrin Glue” applied to surgical sites.
Cryoprecipitate – volume 15ccs
Cryoprecipitate-reduced Plasma
• Cryoprecipitate-reduced plasma is used exclusively for
plasma exchange or transfusion in patients with
thrombotic thrombocytopenic purpura.
• Cryoprecipitate-reduced plasma is the residual
component fluid of FFP from which the cryoprecipitate
has been removed, and therefore contains decreased
amounts of vWF, factor VII, factor XIII and fibrinogen.
• Cryoprecipitate-reduced plasma is stored at −18°C with
an expiration date of one year, and once thawed at 30–
37°C is stored at 1–6°C for up to five days.
Plasma Derivatives
• Plasma derivatives are prepared by cold
ethanol fractionation of large pools of source
or recovered plasma, and include albumin,
immune globulin (Rh immune globulin and
intravenous immunoglobulin), and
coagulation factor concentrates (factor VIII,
factor IX and antithrombin III).
Preparation of platelet concentrate
RBCs PRP
Plasma
Platelet
concentrate
Platelet concentrates
• Important in maintaining hemostasis by preventing spontaneous
bleeding or stopping established bleeding in thrombocytopenic
patients via forming a platelet plug (primary hemostasis).
• Prepared from a single unit of whole blood by cytapheresis
(thrombocytapheresis) or by separating PRP from a unit of WB
within 8H of collection and recentrifuged;
• 40-60 mL of plasma is thereafter expelled into another satellite
bag after the re-centrifugation and the remaining bag contains
platelet concentrate.
• It is the most likely component to be contaminated with bacteria
due to their storage at room temperature.
Platelet concentrates
• Each unit of platelet should elevate the platelet count by 5-10,000
μL in a 165lbs/75kg person.
• The expiration is 5 days as a single unit, 4 hours if pooled; and is
best stored at 20-24˚C (RT) with constant agitation.
• Indicated for use as prophylaxis, in dilutional thrombocytopenia
active bleeding due to thrombocytopenia/thrombocytopathy.
• Therapeutic dose for adults is 6 to 10 units and some patients
become "refractory" to platelet therapy.
• An Rh “D" negative patient should be transfused with Rh “D"
negative platelets due to the presence of a small number of RBCs.
Platelet Components
• Platelet products are used for prophylaxis, or
treatment of bleeding secondary to
thrombocytopenia, or dysfunctional platelets.
• Platelets are prepared from whole blood (also
known as whole blood derived, platelet
concentrates, or random donor platelets) or via
automated apheresis (also known as apheresis
derived platelets, ‘platelets,pheresis’ [said
platelets comma pheresis], plateletpheresis, or
single donor platelets).
• Whole blood derived platelets have a volume of
~40–70 ml and must contain >5.5 × 1010
platelets.
• The usual adult dose of whole blood derived
platelets is a pool of four to six concentrates.
• Whole blood derived platelets can be pooled
prior to issue and subsequently tested for
bacterial contamination; a prepooled,
leukoreduced and bacterial contamination tested
product is also approved.
• Apheresis platelets have a volume of ~300 ml and
must contain >3.0 × 1011 platelets in 75% of the
products tested.
• Most apheresis systems use ACD-A as the
anticoagulant-preservative solution.
• The majority of apheresis devices provide products
that are leukoreduced to a residual WBC count of <5
× 106 white blood cells (termed process
leukoreduction) per product and have very low RBC
contamination which is most relevant for transfusion
of D-positive products to D-negative women.
• Apheresis platelets can also be collected into a
platelet additive solution (PAS).
• In the US, one PAS is FDA approved (Inersol,
Fenwal), although several are approved and in
use outside of the US.
• The current generation of PASs replace 65% of
plasma and are composed of a combination of
citrate, phosphate, acetate, magnesium,
potassium, gluconate and/or glucose.
• Platelets stored in PAS as opposed to plasma are
associated with fewer allergic reactions.
• Newer PAS under study replace 95% of plasma.
• Platelets are stored at room temperature (20–
24°C) with gentle agitation for a maximum of five
days.
• Once the system has been opened or the
platelets pooled, the product must be transfused
within four hours.
Buffy Coat Platelets
• Outside of the US, platelets can be prepared from
whole blood using a buffy coat method, which
differs from the US method (typically referred to
as the platelet-rich plasma [PRP] method).
• Instead of removing the platelet-rich plasma after
softly centrifuging whole blood followed by
separation of platelets from plasma using a hard
spin (the PRP method), the whole blood first goes
through a hard spin, after which the RBCs and
plasma are removed above and below the buffy
coat.
• The buffy coat is then softly spun, after which
the residual white blood cells are removed,
leaving the platelet concentrate.
• Four to six buffy coat platelet concentrates are
then pooled and re-suspended in one of the
donor’s plasma or in PAS to create a single
pooled platelet product.
• Studies have shown no difference in platelet
quality between these two methods of
preparing platelet concentrates; however, the
buffy coat method has several advantages,
including a decrease in residual WBC, and
possible automation of platelet preparation
from whole blood.
Platelet Modification
• Platelet products can be further modified for
specific patients’ needs.
• Multiple modifications may be performed on a
single product.
• Examples of product modifications and their
use include the following:
• Volume-reduction: Platelet products are
volume-reduced to remove the supernatant.
• The primary indications are to prevent
hemolytic reactions by removing the plasma
in ABO-incompatible products, allergic
reactions or volume overload in at-risk
patients.
• Washing: Platelet products are washed to remove
plasma proteins, in order to prevent recurrence of
severe allergic/anaphylactic reactions.
• Leukoreduction: Leukoreduced platelets are used
to mitigate febrile non-hemolytic transfusion
reactions, human leukocyte antigen (HLA)
alloimmunization, Cytomegalovirus (CMV)
transmission, and transfusion-related
immunomodulation.
• Irradiation: Irradiated platelets are used to
prevent transfusion-associated graft versus
host disease.
• Aliquots: Apheresis platelet products can be
divided into smaller volumes for neonatal
transfusions.
Granulocyte concentretes
• Granulocytes, administered for treatment of
patients with neutropenia and life threatening
infection, have a 24-hour shelf-life.
• The use of granulocytes varies due to issues with
rendering timely viral testing and in transporting
of the product from collection to transfusion
service during that timeframe, and the
presumption of the collection of inadequate
doses of granulocytes using currently accepted
methods of donor preparation and collection.
• Granulocytes are usually collected using
apheresis devices a process called
leukocytapheresis, but they may also be
prepared from the buffy coat of centrifuged
whole blood for neonatal transfusions.
• Apheresis-derived granulocytes have a volume
of ~200 ml and contain RBCs (10–50 ml),
platelets (~3 × 1011) and plasma in addition to
granulocytes (>1 × 1010).
• Modalities used to improve the granulocyte
dose are to stimulate the donor with
granulocyte colony stimulating factor (G-CSF)
and/or corticosteroids, although this method
and its resultant clinical efficacy have not as
yet been proven in randomized clinical trials,
and/or to use hydroxyethyl starch (HES) to
improve sedimentation and collection during
apheresis.
• The use of all three modalities should increase
the product yield to >1 × 1011 granulocytes.
• These products should be transfused as soon as
possible, but they may be stored for up to 24
hours after collection at 20–24°C without
agitation.
• All granulocyte products should be irradiated to
prevent transfusion-associated graft versus host
disease, but they cannot be leukoreduced and,
therefore, CMV-seronegative products may be
indicated for at-risk patients.
Massive Transfusion
• 10 or more pRBC units (TBV) in <24
hours.
• Others:
– Replacement of 50% of TBV within 3 hours.
– Blood loss >150 ml/min.
Massive Transfusion
Clinical Settings
• Trauma
• Surgery (e.g. Liver, Cardiovascular)
• GI bleeding
• Obstetrics
Laboratory Values to Monitor in
Trauma
• Hgb/Hct
• INR/PTT
• Fibrinogen
• Electrolytes
• Platelet Count
• Blood Gases
• FFP if INR> 1.5 or PT >1.5 X Normal
• Platelets if Count <50K-100K
• Cryoprecipitate if Fibrinogen <100mg/dl
(each unit contains ~250 mg)
Indications for Platelets & Hemostatic
Factors
Massive Exanguination
“Triad of Death”
• Acidosis
• Hypothermia
• Coagulopathy
Massive Transfusion Protocol
• Mortality in massive transfusion is high –
up to 57% (patients transfused >50 RBC
units)
• Coagulopathy is present early and not only
a factor of hemodilution (Gonzalez et al
2007)
• A recent retrospective review shows an
increase in survival with a 1:1:1 ratio of
plasma: platelets: RBCs
Massive Transfusion Protocol
• New Trend to give RBCs, FFP and
Platelets to simulate whole blood
• Typical Published Ratios of
RBC:FFP:Platelets using Typical Products
– 6 units RBC Adult (250ml/unit)
– 6 units FFP (~250ml/unit)
– 6 units Platelet Concentrate (50ml/unit)
Massive Transfusion Guideline
(Established 2008)
• Adult Replacement Volumes established
based on Acute Blood Loss of 50% and
maintenance of a RBC:FFP:Platelet ratio of
whole blood AND using the products available
at UCDMC
– 6 units RBC Adult (250ml/unit)
– 3 units FFP Jumbo (400ml/unit)
– 1 unit Plateletpheresis (250ml/unit)
Potential Adverse Effects of Massive
Transfusion
• Metabolic Disturbances
• Transfusion Reactions
• Infectious Disease Risks
• Citrate Toxicity
• Hyperkalemia
• Decreased Oxygen Delivery
• Hypothermia
Acute/Immediate Transfusion
Reactions
• Acute Hemolytic Reactions
• Bacterial Contamination of Blood Products
• Anaphylaxis
• Transfusion Related Acute Lung Injury
• Severe Febrile Reactions
• Transfusion Associated Circulatory Overload
• Metabolic Problems of Massive Transfusion
• Air Emboli & Microemboli
• Hypotensive Response to Plasma
REFERENCE
• Transfusion Medicine and Hemostasis Clinical and Laboratory
Aspects second edition by Beth H. Shaz, C.D. Hillyer, M. Roshal, C.S.
Abrams.
• http://Portal.abuad.edu.ng> …PPT mls 306 preparation of blood
and blood components therapy accessed on 23 july 2021
• Basu D, Kulkarni R. Overview of blood components and their
preparation. Indian J Anaesth 2014;58:529-37 accessed on 23 july
2021
• Gonzalez EA, Moore FA, Holcomb JB, et al. (2007) Fresh frozen
plasma should be given earlier to patients requiring massive
transfusion. The Journal of Trauma, 62: 112-119.
•

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3.0 blood components introduction s.pptx

  • 1. BLOOD COMPONENT PREPARATION, STORAGE AND USE BY Salvatory Temu 18/12/2023
  • 2. BLOOD COMPONENTS • Blood components, including packed red blood cell (PRBC) concentrate, fresh frozen plasma and cryoprecipitate, platelet concentrate and granulocyte products. • Prepared either from whole blood donation or by automated apheresis. • Blood component preparation and manufacture allow each component to be manufactured and stored under optimal conditions.
  • 4. • Apheresis is a procedure where required single or more than one component is collected, and the rest of blood components are returned back to the donor. • The working principle of apheresis equipment is either by centrifugation (different specific gravity) or by filtration (different size). BLOOD COMPONENTS
  • 5. • The use of components (i.e. component therapy) has largely replaced the use of whole blood due to the ability to choose individual components and component constituents that target a specific patient’s needs as well as enabling each component to be stored optimally. BLOOD COMPONENTS
  • 6. • The blood components being foreign to a patient may produce adverse effects that may range from mild allergic manifestations to fatal reactions. • Such reactions are usually caused by plasma proteins, leucocytes, red cell antigens, plasma and other pathogens. • To avoid and reduce such complications, blood products are modified as leukoreduced products, irradiated products, volume reduced products, saline washed products and pathogen inactivated products. BLOOD COMPONENTS
  • 7. Primary blood bag with satellite bags BLOOD COMPONENTS
  • 8. Key terminologies in blood preparation • Whole blood: blood collected before separation into components. • Components: parts of whole blood that are separated. • Closed system: a sterile system of blood collection. • Open system: when the collection is exposed to air, decreasing expiration date.
  • 9. Collection of Blood and blood components • Blood is collected into a primary bag containing anticoagulant- preservatives and depending on the components choice; satellite bags with hermetic seal in between may also be attached. • The blood bag anticoagulant-preservative minimizes biochemical changes and increases shelf life of the component or whole blood. • Blood unit is collected into a closed system within 15 minutes to avoid activating the coagulation system. • Transfuse within 24 hours if the hermetic seal is broken and stored at 1-4˚C, or within 4 hours if stored at 20-24˚C.
  • 10. Whole Blood (WB) • Clinical indications for use of WB are now becoming extremely limited. Whole blood comprises RBCs, WBCs, platelets and plasma (with anticoagulant). • Used for massive transfusion to correct acute hypovolaemia such as trauma and shock, exchange transfusion. • 1 unit increases Hgb 1 g/dL and Hct 3% • RARELY used today, platelets non-functional, labile coagulation factors gone. • Must be ABO identical.
  • 11. Whole Blood • Whole blood is the most common starting product for component preparation and manufacture. • Whole blood is generally not found in most blood banks today because component therapy is more appropriate to target a patient’s specific indications for transfusion (for example, RBC products for symptomatic anemia, plasma products for multiple coagulation factor deficiencies, and platelet products for thrombocytopenia or platelet dysfunction).
  • 12. • In addition, in whole blood, platelet function and post-transfusion survivability are lost when the whole blood is stored in the refrigerator, and coagulation factor activity, especially for labile factors (factors V and VIII), deteriorates over time.
  • 13. • In general, 500 ml of whole blood is collected into a bag with 70 ml of anticoagulant- preservative solution (see below), creating a product with a final hematocrit of ~38%. • Whole blood is stored at 1–6°C. • Depending on the manufacturer’s system and a country’s regulations, different hold times are allowed before the preparation of blood components from whole blood.
  • 14. • If platelet products are to be manufactured from whole blood, the whole blood product must be stored at room temperature until the platelets are removed.
  • 15. • Fresh whole blood is occasionally used by the US military via ‘walking blood donors’ (donors who are ABO/D typed, infectious disease tested and available to donate in times of need).
  • 16. • This product is transfused as soon as possible and has a shelf-life of 24 hours when stored at room temperature, allowing maintenance of platelet function, while still minimizing the risk of bacterial overgrowth.
  • 17. • The primary use of this product is to infuse viable platelets when other platelet products are not available. • Risks associated with use of this product include transfusion-transmitted diseases and transfusion associated graft versus host disease.
  • 18. • Blood component preparation was developed in 1960 to separate blood products from one unit whole blood by a specialised equipment called as refrigerated centrifuge.
  • 19. Component Manufacturing • When whole blood is manufactured into components, it is collected into a primary bag containing an anticoagulant-preservative solution. • The primary bag has up to three attached satellite bags for RBC, platelet, and plasma or cryoprecipitate component manufacturing.
  • 20. Diagrammatic representation of blood components preparation
  • 21. • Due to the different specific gravities of RBCs (1.08–1.09), plasma (1.03–1.04) and platelets (1.023), differential centrifugation of the whole blood unit is used to prepare blood components. • Optimal component separation requires specific centrifugation variables, such as rotor size, speed, and duration of spin (Figure 9.1).
  • 22.
  • 23. • Preparing only PRBC and fresh frozen plasma(FFP) is by single-step heavy spin centrifugation; however preparing platelet concentrates(PLTCs), PRBC concentrates and FFP is by two step centrifugation.
  • 24. Anticoagulant-Preservative Solutions • Whole blood is collected into containers with anticoagulant-preservative solutions, including acid-citrate-dextrose (ACD), citrate-phosphate- dextrose (CPD), citrate-phosphate-dextrose- dextrose (CP2D) and citrate-phosphate-dextrose- adenine (CPDA-1). • The citrate (sodium citrate and citric acid) acts as an anticoagulant, and the phosphate (monobasic sodium phosphate and trisodium phosphate), adenine and dextrose are substrates for cellular metabolism.
  • 25. Packed RBC Components • RBC products are used primarily for the treatment of symptomatic anemia or hemorrhage in order to increase tissue oxygenation. • RBC products are prepared either by centrifugation of whole blood followed by removal of 200- 250ml of platelet-rich plasma layer, or by automated apheresis collection. • Anticoagulant preservative solutions allow RBC components to be stored for extended periods of time at 1–6°C without a significant detrimental effect on the quality of the RBC.
  • 26. • Prepared by sedimenting whole blood (WB) or centrifuging it to remove 200-250ml of supernatant plasma from a unit of WB. • This produces 200-250ml red cell concentrates with a haematocrit of approximately 80% for non-additive (CPD), 60% for additive (ADSOL). • A unit increases patient’s Hb level by about 1g/dL (10g/L) and haematocrit by 3%. • It does not contain functional platelets or granulocytes and has same O2 carrying capacity with W.B. • It is used to treat symptomatic anaemia and routine blood loss during surgery to increase patient’s red cell mass without increasing their blood volume.
  • 27. • The shelf-life of a given product is determined by the solution used and based on the criterion that there is less than 1% hemolysis in the product at the end of storage, and 75% of the transfused RBCs remain viable 24 hours after transfusion. • RBCs stored in CPD or CP2D have a shelf-life of 21 days and in CPDA-1 of 35 days.
  • 28. • RBCs stored in CPD, CP2D or CPDA-1 have a hematocrit of ~80% with a final volume of 225–350 ml and up to 110 ml of plasma. • The majority of RBC products are stored in additive solutions, which extend the shelf-life of the product to 42 days.
  • 29. Packed red cell concentrate: Once the unit of red cell concentrate is “opened”, it has a 24 hour expiration date.
  • 30. Additive Solutions • Additive solutions, such as AS-1, AS-3 and AS- 5, can be added to the primary anticoagulant- preservative solution to increase the shelf-life of RBCs to 42 days. • All of these solutions contain dextrose, adenine and sodium chloride; AS-1 and AS-5 contain mannitol, and AS-3 contains monobasic sodium phosphate, sodium citrate and citric acid.
  • 31. • Additive solution products have a final volume of 350– 400 ml and hematocrit of 55–65%, containing 100–110 ml of additive solution and 10–40 ml of plasma. • In general, additive solutions must be added to the product within 24 hours of collection, depending on manufacturer’s specifications. • Outside of the US, SAGM (saline-adenine-glucose- mannitol), which is hypertonic, is used and PAGGSM (phosphate-adenine-glucose-guanosin-saline- mannitol), which is isotonic, is under development.
  • 32. • A number of other additive solutions are currently under study, including EAS-81 (NaHCO3, Na2HPO4, adenine, dextrose, and mannitol) as well as methods to decrease oxidative damage by adding GSH precursors (glutamine, glycine and N-acetyl-L-cysteine) or O2 removal.
  • 33. RBC Modification • RBC products can be further modified for specific patients’ needs. • Multiple modifications can be performed on a single product. • Examples of product modifications and their use include the following:
  • 34. • Freezing: Frozen RBC products, which are cryopreserved with glycerol, have an extended storage life of up to 10 years when stored at −65°C or below. • The primary indications for frozen RBCs are for rare RBC phenotypes (e.g. products from donors who lack high frequency RBC antigens) or autologous donations, or to maintain product inventory for disaster management. • Frozen RBC products must be deglycerolized prior to transfusion. • Deglycerolized products have a 24-hour outdate.
  • 35. • Volume reduction: RBC products can be volume- reduced to remove most of the supernatant. • The primary indications are to remove the accumulated potassium, which leaks from the RBCs during storage, in order to prevent hyperkalemia in at-risk patients such as neonates; to reduce the AS concentration, though this is rarely required or appropriate; or to reduce the volume infused in volume-sensitive patients.
  • 36. • Washing: RBC products can be washed to remove the plasma and, with it, plasma proteins. • This process can be used to minimize the risk of recurrence of severe allergic/anaphylactic reactions.
  • 37. • Rejuvenation: Rejuvenated RBCs have 2,3-DPG and ATP levels near to those of a freshly drawn unit. • During storage 2,3-DPG levels fall linearly towards zero during the first two weeks. • ATP levels fall to 50–70% of initial levels during storage, depending on the anticoagulant- preservative solution used.
  • 38. • Rejuvenation is usually performed prior to freezing RBCs. • The RBCs must be washed prior to use to remove the rejuvenation solution.
  • 39. • Leukoreduction: Leukoreduced RBCs are used to mitigate the risk of febrile nonhemolytic transfusion reactions, HLA alloimmunization, CMV transmission and transfusion-related immunomodulation, each in appropriate and clinically indicated populations. • Most institutions and clinicians prefer universally available leukoreduced products.
  • 40. • Irradiation: Irradiated RBCs are used to prevent transfusion-associated graft versus host disease. • Aliquots: RBC products can be divided into smaller volumes for neonatal transfusions.
  • 41. Plasma Components • Plasma components are primarily used to treat multiple plasma factor deficiencies or to prevent development of a coagulopathy. • Plasma contains all of the coagulation factors found in whole blood and the critical proteins albumin and fibrinogen. • Plasma products may be manufactured either from whole blood or by automated apheresis. • Plasma products that are used in the transfusion service include fresh frozen plasma (FFP), plasma frozen within 24 hours of phlebotomy (FP24) and thawed plasma (TP).
  • 42. • FFP must be frozen within 6–8 hours of collection, while FP24 is frozen between 8 and 24 hours of collection. • FP24 has lower levels of factors V and VIII than FFP, but this reduction is rarely of significance. • FFP and FP24 are stored at −18°C or colder for up to one year (with approval from the FDA, FFP can be stored at −65°C or colder for up to seven years). • Prior to transfusion, plasma must be thawed at 30– 37°C, which requires ~20 minutes; it is then stored at 1–6°C if not transfused immediately.
  • 43. • Once thawed, FFP and FP24 must be transfused within 24 hours, otherwise it becomes classified as TP, which may be stored at 1–6°C for up to five days. • During these five days of storage, factor VIII activity levels decrease by approximately 30%. • FFP, FP24 and TP can be used interchangeably in most situations.
  • 44. • Plasma products may be further manufactured into cryoprecipitate and cryoprecipitate-reduced plasma. • In addition, outside of the US, large pools of plasma are manufactured into solvent/detergent-treated plasma, which has minimal (if any) risk of transmitting lipid- enveloped viruses (HIV, HCV, and HBV).
  • 45. • Other pathogen inactivated plasma products available outside of the US, which are not pooled, include methylene blue (MB), riboflavin and ultraviolet light, and amotosalen and ultraviolet light though some manufacturers and countries have considered removing MB from use due to allergic reactions.
  • 46. • Plasma (stored at −18°C or below) and liquid plasma (stored at 1–6°C) are manufactured from whole blood no later than five days after the whole blood expiration date, and are converted into an unlicensed product termed ‘recovered plasma’ that can be shipped for further fractionation into albumin, antithrombin III, factor VIII or factor IX concentrates, or immunoglobulin preparations. • Liquid plasma is sometimes, though rarely, used for transfusion.
  • 47. Fresh Frozen Plasma • Prepared by removing plasma from WB within 8 hours of collection and must be frozen within same duration. • It is used to replace labile and non-labile coagulation factors in massively bleeding patients and treat bleeding associated with clotting factor deficiencies when factor concentrate is not available. • Its constituents are water, carbohydrates, fats, minerals, proteins (all labile and stable clotting factors). • Each unit of FFP measures 200-225ml and elevates the level of each clotting factor by 2-3% in adults, the therapeutic dose is 10-15ml/kg.
  • 48. Fresh Frozen Plasma • The storage temperatures: - frozen -18˚C, preferably -30˚C or lower - thawed - 1-6˚C - Thawed in 30-37˚C water bath. • Expiration: - frozen - 1 year if stored at <-18˚C. - frozen - 7 years if stored at <-65˚C. - thawed - 24 hours. • Must have mechanism to detect units which have thawed and refrozen due to improper storage. • Must be ABO compatible.
  • 49. • FFP is thawed before transfusion at 30-37°C in a waterbath for 30-45 minutes and can then be stored at 1-6°C and transfused within 24 hours. Fresh Frozen Plasma
  • 50. Fresh Frozen Plasma • It is indicated for use in the following conditions: - Replacement for isolated coagulation function deficiencies. - The reversal of warfarin therapies. - In the case of massive blood transfusion. - Antithrombin III deficiency treatment. - Correction of coagulopathy or liver disease. - Thrombotic thrombocytopenic purpura.
  • 51. Cryoprecipitated antihemophilic factor (AHF) • The primary indication for the use of cryoprecipitate (the FDA term is cryoprecipitated AHF) is for replacement of fibrinogen. • Cryoprecipitate is prepared by slowly thawing FFP to 1–6°C, thereby leading to formation of a precipitate, which is collected and refrozen to make cryoprecipitate. • Each unit of cryoprecipitate, which is approximately 15 ml, must contain fibrinogen (>150 mg) and factor VIII (>80 IU) at these specified levels.
  • 52. • In addition, each unit contains von Willebrand’s factor (vWF) (80– 120 IU), factor XIII (40–60 IU) and fibronectin. • Cryoprecipitate is usually administered to adults in doses of 8–10 donor units, which are pooled prior to transfusion (known as a ‘cryopool’). • Some blood centers manufacture prepooled cryoprecipitate, where five units are pooled together prior to storage. • This product is easier for the transfusion service to use in cases of emergency than pooling prior to issue (post-thaw). • Cryoprecipitate units can be stored for up to one year at −18°C and must be transfused within four hours of pooling using an open system and within six hours of thawing using a closed system.
  • 53. Cryoprecipitated antihemophilic factor (AHF) • Cryoprecipitated antihemophilic factor (AHF) or “Cryo” is the cold-insoluble portion of plasma that precipitates when FFP is thawed at 1-6˚C. • It contains high levels of Factor VIII (about 80 IU in each unit), plasma (10-15ml), von Willebrand’s factor (platelet adhesion), Fibrinonectin and Fibrinogen (150 mg in each unit). • It is used for treatment of hemophiliacs and Von Willebrand disease when concentrates are unavailable. • Used most commonly for patients with DIC or low fibrinogen levels and the therapeutic dose for an adult is 6 to 10 units.
  • 54. Cryoprecipitated antihemophilic factor (AHF) • Can be prepared from WB which is then designated as "Whole Blood Cryoprecipitate Removed" or from FFP. • The plasma is first frozen, then thawed at 1-6˚C which results in the formation of a precipitate. • The plasma is centrifuged, cryoprecipitate goes to the bottom. • Plasma is then expelled or removed and frozen at -18 ˚C within 1 hour of preparation.
  • 55. FFP Frozen within 8 hours Thawed FFP Cryoprecipitate (VIII, vW) Plasma cryoprecipitate reduced (TTP, FII, V, Vii, IX, X, XI) Thaw at 30-37°C Store at RT 4 hrs Refrozen with 24 hrs of separation Store at ≤18°C 1 yr 5 day expiration at 1-6°C
  • 56. Cryoprecipitated antihemophilic factor (AHF) • Same storage as FFP (cannot be re-frozen as FFP once it is separated); at -18˚C for 1 year and if thawed, store at room temperature for 4 hours. • The leftover plasma is called “cryoprecipitate-reduced” or "plasma cryo". • Cryoprecipitate-reduced used for thrombotic thrombocytopenic purpura (TTP), 2° treatment for Factor VIII deficiency (Hemophilia A), 2 ° treatment for von Willebrand’s Disease, Congenital or acquired fibrinogen deficiency, FXIII deficiency and “Fibrin Glue” applied to surgical sites.
  • 58. Cryoprecipitate-reduced Plasma • Cryoprecipitate-reduced plasma is used exclusively for plasma exchange or transfusion in patients with thrombotic thrombocytopenic purpura. • Cryoprecipitate-reduced plasma is the residual component fluid of FFP from which the cryoprecipitate has been removed, and therefore contains decreased amounts of vWF, factor VII, factor XIII and fibrinogen. • Cryoprecipitate-reduced plasma is stored at −18°C with an expiration date of one year, and once thawed at 30– 37°C is stored at 1–6°C for up to five days.
  • 59. Plasma Derivatives • Plasma derivatives are prepared by cold ethanol fractionation of large pools of source or recovered plasma, and include albumin, immune globulin (Rh immune globulin and intravenous immunoglobulin), and coagulation factor concentrates (factor VIII, factor IX and antithrombin III).
  • 60. Preparation of platelet concentrate RBCs PRP Plasma Platelet concentrate
  • 61. Platelet concentrates • Important in maintaining hemostasis by preventing spontaneous bleeding or stopping established bleeding in thrombocytopenic patients via forming a platelet plug (primary hemostasis). • Prepared from a single unit of whole blood by cytapheresis (thrombocytapheresis) or by separating PRP from a unit of WB within 8H of collection and recentrifuged; • 40-60 mL of plasma is thereafter expelled into another satellite bag after the re-centrifugation and the remaining bag contains platelet concentrate. • It is the most likely component to be contaminated with bacteria due to their storage at room temperature.
  • 62. Platelet concentrates • Each unit of platelet should elevate the platelet count by 5-10,000 μL in a 165lbs/75kg person. • The expiration is 5 days as a single unit, 4 hours if pooled; and is best stored at 20-24˚C (RT) with constant agitation. • Indicated for use as prophylaxis, in dilutional thrombocytopenia active bleeding due to thrombocytopenia/thrombocytopathy. • Therapeutic dose for adults is 6 to 10 units and some patients become "refractory" to platelet therapy. • An Rh “D" negative patient should be transfused with Rh “D" negative platelets due to the presence of a small number of RBCs.
  • 63. Platelet Components • Platelet products are used for prophylaxis, or treatment of bleeding secondary to thrombocytopenia, or dysfunctional platelets. • Platelets are prepared from whole blood (also known as whole blood derived, platelet concentrates, or random donor platelets) or via automated apheresis (also known as apheresis derived platelets, ‘platelets,pheresis’ [said platelets comma pheresis], plateletpheresis, or single donor platelets).
  • 64. • Whole blood derived platelets have a volume of ~40–70 ml and must contain >5.5 × 1010 platelets. • The usual adult dose of whole blood derived platelets is a pool of four to six concentrates. • Whole blood derived platelets can be pooled prior to issue and subsequently tested for bacterial contamination; a prepooled, leukoreduced and bacterial contamination tested product is also approved.
  • 65. • Apheresis platelets have a volume of ~300 ml and must contain >3.0 × 1011 platelets in 75% of the products tested. • Most apheresis systems use ACD-A as the anticoagulant-preservative solution. • The majority of apheresis devices provide products that are leukoreduced to a residual WBC count of <5 × 106 white blood cells (termed process leukoreduction) per product and have very low RBC contamination which is most relevant for transfusion of D-positive products to D-negative women.
  • 66. • Apheresis platelets can also be collected into a platelet additive solution (PAS). • In the US, one PAS is FDA approved (Inersol, Fenwal), although several are approved and in use outside of the US. • The current generation of PASs replace 65% of plasma and are composed of a combination of citrate, phosphate, acetate, magnesium, potassium, gluconate and/or glucose.
  • 67. • Platelets stored in PAS as opposed to plasma are associated with fewer allergic reactions. • Newer PAS under study replace 95% of plasma. • Platelets are stored at room temperature (20– 24°C) with gentle agitation for a maximum of five days. • Once the system has been opened or the platelets pooled, the product must be transfused within four hours.
  • 68. Buffy Coat Platelets • Outside of the US, platelets can be prepared from whole blood using a buffy coat method, which differs from the US method (typically referred to as the platelet-rich plasma [PRP] method). • Instead of removing the platelet-rich plasma after softly centrifuging whole blood followed by separation of platelets from plasma using a hard spin (the PRP method), the whole blood first goes through a hard spin, after which the RBCs and plasma are removed above and below the buffy coat.
  • 69. • The buffy coat is then softly spun, after which the residual white blood cells are removed, leaving the platelet concentrate. • Four to six buffy coat platelet concentrates are then pooled and re-suspended in one of the donor’s plasma or in PAS to create a single pooled platelet product.
  • 70. • Studies have shown no difference in platelet quality between these two methods of preparing platelet concentrates; however, the buffy coat method has several advantages, including a decrease in residual WBC, and possible automation of platelet preparation from whole blood.
  • 71. Platelet Modification • Platelet products can be further modified for specific patients’ needs. • Multiple modifications may be performed on a single product. • Examples of product modifications and their use include the following:
  • 72. • Volume-reduction: Platelet products are volume-reduced to remove the supernatant. • The primary indications are to prevent hemolytic reactions by removing the plasma in ABO-incompatible products, allergic reactions or volume overload in at-risk patients.
  • 73. • Washing: Platelet products are washed to remove plasma proteins, in order to prevent recurrence of severe allergic/anaphylactic reactions. • Leukoreduction: Leukoreduced platelets are used to mitigate febrile non-hemolytic transfusion reactions, human leukocyte antigen (HLA) alloimmunization, Cytomegalovirus (CMV) transmission, and transfusion-related immunomodulation.
  • 74. • Irradiation: Irradiated platelets are used to prevent transfusion-associated graft versus host disease. • Aliquots: Apheresis platelet products can be divided into smaller volumes for neonatal transfusions.
  • 75. Granulocyte concentretes • Granulocytes, administered for treatment of patients with neutropenia and life threatening infection, have a 24-hour shelf-life. • The use of granulocytes varies due to issues with rendering timely viral testing and in transporting of the product from collection to transfusion service during that timeframe, and the presumption of the collection of inadequate doses of granulocytes using currently accepted methods of donor preparation and collection.
  • 76. • Granulocytes are usually collected using apheresis devices a process called leukocytapheresis, but they may also be prepared from the buffy coat of centrifuged whole blood for neonatal transfusions. • Apheresis-derived granulocytes have a volume of ~200 ml and contain RBCs (10–50 ml), platelets (~3 × 1011) and plasma in addition to granulocytes (>1 × 1010).
  • 77. • Modalities used to improve the granulocyte dose are to stimulate the donor with granulocyte colony stimulating factor (G-CSF) and/or corticosteroids, although this method and its resultant clinical efficacy have not as yet been proven in randomized clinical trials, and/or to use hydroxyethyl starch (HES) to improve sedimentation and collection during apheresis.
  • 78. • The use of all three modalities should increase the product yield to >1 × 1011 granulocytes. • These products should be transfused as soon as possible, but they may be stored for up to 24 hours after collection at 20–24°C without agitation. • All granulocyte products should be irradiated to prevent transfusion-associated graft versus host disease, but they cannot be leukoreduced and, therefore, CMV-seronegative products may be indicated for at-risk patients.
  • 79. Massive Transfusion • 10 or more pRBC units (TBV) in <24 hours. • Others: – Replacement of 50% of TBV within 3 hours. – Blood loss >150 ml/min.
  • 80. Massive Transfusion Clinical Settings • Trauma • Surgery (e.g. Liver, Cardiovascular) • GI bleeding • Obstetrics
  • 81. Laboratory Values to Monitor in Trauma • Hgb/Hct • INR/PTT • Fibrinogen • Electrolytes • Platelet Count • Blood Gases
  • 82. • FFP if INR> 1.5 or PT >1.5 X Normal • Platelets if Count <50K-100K • Cryoprecipitate if Fibrinogen <100mg/dl (each unit contains ~250 mg) Indications for Platelets & Hemostatic Factors
  • 83. Massive Exanguination “Triad of Death” • Acidosis • Hypothermia • Coagulopathy
  • 84. Massive Transfusion Protocol • Mortality in massive transfusion is high – up to 57% (patients transfused >50 RBC units) • Coagulopathy is present early and not only a factor of hemodilution (Gonzalez et al 2007) • A recent retrospective review shows an increase in survival with a 1:1:1 ratio of plasma: platelets: RBCs
  • 85. Massive Transfusion Protocol • New Trend to give RBCs, FFP and Platelets to simulate whole blood • Typical Published Ratios of RBC:FFP:Platelets using Typical Products – 6 units RBC Adult (250ml/unit) – 6 units FFP (~250ml/unit) – 6 units Platelet Concentrate (50ml/unit)
  • 86. Massive Transfusion Guideline (Established 2008) • Adult Replacement Volumes established based on Acute Blood Loss of 50% and maintenance of a RBC:FFP:Platelet ratio of whole blood AND using the products available at UCDMC – 6 units RBC Adult (250ml/unit) – 3 units FFP Jumbo (400ml/unit) – 1 unit Plateletpheresis (250ml/unit)
  • 87. Potential Adverse Effects of Massive Transfusion • Metabolic Disturbances • Transfusion Reactions • Infectious Disease Risks • Citrate Toxicity • Hyperkalemia • Decreased Oxygen Delivery • Hypothermia
  • 88. Acute/Immediate Transfusion Reactions • Acute Hemolytic Reactions • Bacterial Contamination of Blood Products • Anaphylaxis • Transfusion Related Acute Lung Injury • Severe Febrile Reactions • Transfusion Associated Circulatory Overload • Metabolic Problems of Massive Transfusion • Air Emboli & Microemboli • Hypotensive Response to Plasma
  • 89. REFERENCE • Transfusion Medicine and Hemostasis Clinical and Laboratory Aspects second edition by Beth H. Shaz, C.D. Hillyer, M. Roshal, C.S. Abrams. • http://Portal.abuad.edu.ng> …PPT mls 306 preparation of blood and blood components therapy accessed on 23 july 2021 • Basu D, Kulkarni R. Overview of blood components and their preparation. Indian J Anaesth 2014;58:529-37 accessed on 23 july 2021 • Gonzalez EA, Moore FA, Holcomb JB, et al. (2007) Fresh frozen plasma should be given earlier to patients requiring massive transfusion. The Journal of Trauma, 62: 112-119. •