Preparation of Red Blood Cells

Canadian Blood Services produces three types of Red Blood Cells (RBC):

• AS-3 RBCs, LR (Leukocytes reduced)
• CP2D RBCs, LR (Leukocytes reduced)
• CPDA-1 RBCs, LR (Leukocytes reduced)

Following the implementation of the Buffy Coat Production Method (BCPM), a fourth red blood cell product will also be available:

• SAGM RBCs, LR (Leukocytes reduced)

Preparation of AS-3 RBC

Below is a description of how AS-3 RBCs are prepared from a whole blood collection. Leukofiltration is not shown, but is done prior to centrifugation in Step 1.

Step 1		The whole blood donation is centrifuged to separate red cells from plasma.
Step 2		Approximately 190-260 mL of donor plasma is expressed into the first satellite bag (the maximum amount of plasma is removed) The plasma can be used to prepare Fresh Frozen Plasma (FFP), Frozen Plasma (FP) or cryoprecipitate and cryosupernatant plasma. Alternatively, the plasma may be shipped as recovered plasma for further manufacture.
Step 3		The AS-3 (Nutricel ™) is added to the RBC
Step 4		The AS-3 RBC LR unit is sealed and stored at 1-6oC for 42 days.

Preparation of CP2D or CPDA-1 RBC

Step 1		The whole blood donation is centrifuged to separate red cells from plasma.
Step 2		Approximately 190-260 mL of donor plasma is expressed into the satellite bag. The plasma can be used to prepare FFP, FP or cryoprecipitate and cryosupernatant plasma. The plasma may also be shipped as recovered plasma for further manufacture.
Step 3		The RBC unit is sealed and stored at 1-6°C. CP2D RBCs have a shelf life of 21 days. CPDA-1 RBCs may be stored for 35 days from

Additional information on RBCs may be found in the Circular of Information for the Use of Human Blood and Blood Components - 2004.

Preparation of SAGM RBC

When RBCs are prepared by the BCPM, the whole blood donation is centrifuged using a “hard spin” to separate red cells from plasma  (the buffy coat layer containing the platelets appears as a thick white layer above the red cells). Using the component separator (Compomat G4) RBCs and plasma are extracted from the buffy coat (from the bottom and top of the primary bag respectively). See preparation of random donor platelets for additional information.

Below is a simplified explanation of how SAGM Red Blood Cells (LR) is prepared from a whole blood collection using the wholed blood filtration method.  Leukofiltration is not shown.

Step 1		The whole blood donation is centrifuged to separate red cells from plasma. Segments are prepared and secured to the blood pack.
Step 2		The plasma is separated from the red blood cells using the Compomat G4 (a semi-automated instrument for extracting the plasma).  The SAGM additive is then added to the red cells.
Step 3		The SAGM RBC unit is gently mixed until homogenous and stored at 1-6° C for a maximum of 42 days.

Additional information on SAGM RBCs (LR), may be found in the Circular of Information for the Use of Human Blood and Blood Components - 2005.

Preparation of Plasma Products

Canadian Blood Services manufactures the following plasma products:

• CP2D Fresh Frozen Plasma, LR (Leukocytes reduced)
• CPDA-1 Fresh Frozen Plasma, LR (Leukocytes reduced)
• Fresh Frozen Plasma Apheresis
• Frozen Plasma, LR (Leukocytes reduced)

Following the implementation of the Buffy Coat Production Method (BCPM), another plasma product will also be available:

• CPD Frozen Plasma

For information about cryosupertant plasma, see Preparation of Cryoprecipitate and Cryosupernatant Plasma.

Preparation of CP2D or CPDA-1 Fresh Frozen Plasma (FFP) and Frozen Plasma (FP)

Fresh Frozen Plasma is plasma that is frozen within 8 hours of collection. It may be prepared from a whole blood donation (by separating the red cells and plasma collected) or from an apheresis collection.

Frozen Plasma is plasma that is frozen within 24 hours of collection. All Frozen Plasma prepared by Canadian Blood Services comes from whole blood donations.

Below is a simplified explanation of how these products are prepared from a whole blood collection. Leukofiltration is not shown. Note that Fresh Frozen Plasma can also be prepared when manufacturing platelets.

Step 1		The whole blood donation is centrifuged to separate red cells from plasma.
Step 2		Approximately 190-260 mL of donor plasma is expressed into the first satellite bag. The RBC unit is sealed and stored at 1 – 6oC for its shelf-life. The plasma can be stored as FP or FFP that can be further processed to cryoprecipitate and cryosupernatant plasma using the attached satellite container.
Step 3		Plasma is stored frozen by the manufacturer at temperatures less than minus 20oC for up to 12 months. Non-manufacturers(hospitals and transfusion facilities) are referred to the Canadian Standards Association CAN/CSA-Z902-04, which state that: FFP and FP be stored at -18°C or colder for up to 12 months, and thawed FFP and FP be stored at 1-6°C for up to 24 hours.

Additional information on plasma components may be found in the Circular of Information for the Use of Human Blood and Blood Components - 2004

Preparation of CPD Frozen Plasma

With the implementation of the Buffy Coat Production Method, Frozen Plasma will be prepared from whole blood collected in CPD anticoagulant. Using this production method:

• Plasma will be frozen within 24 hours of collection.
• Frozen plasma from whole blood donations will not be identified as being leukoreduceded, although some will be. Units that are not intended for platelet (buffy coat) production will be leukoreduced as a consequence of whole blood filtration, while those units that are intended for platelet (buffy coat) production will have had the buffy coat removed.

Below is a simplified explanation of how FP from a whole blood collection is prepared. Leukofiltration is not shown.  

Step 1	Whole blood collected into CPD is centrifuged to separate red cells from plasma.
Step 2	Following centrifugation the whole blood is loaded onto the component separator (Compomat G4). The plasma is extracted and frozen within 24 hrs. of collection
Step 3	Plasma is stored frozen by the manufacturer at - 20oC or colder for up to 12 months.   Non-manufacturers (hospitals and transfusion facilities) are referred to the Canadian Standards Association CAN/CSA-Z902-04, which states: Frozen plasma be stored at -18°C or colder for up to12 months, and thawed FP be stored at 1-6°C for up to 24 hours.

 

Preparation of Platelets

Canadian Blood Services produces two platelet components:

• CP2D Platelets, LR (Leukocytes Reduced)
• Platelets, Apheresis, LR (Leukocytes Reduced)

Following the implementation of the Buffy Coat Production Method (BCPM), another platelet product will be available:

• CPD Platelets, Pooled, LR (Leukocytes reduced)

Preparation of CP2D Platelets

CP2D Platelets are prepared from a single unit of whole blood collected in CP2D. This product is also known as random donor platelets.

If platelet preparation is intended, the donor unit must be bled into a specific multiple bag system designed for this purpose. Below is a simple schematic of how random donor platelets are prepared.

The whole blood donation is centrifuged at room temperature to separate red cells from plasma.

Centrifugation at room temperature (20-24° C) is required to prevent the platelets from aggregating. A “light spin” is used to keep the platelets suspended in plasma.
 
Approximately 190-260 mL of donor plasma is expressed through a filter into the first satellite bag to produce platelet-rich plasma (PRP).

The RBC unit and additive pack are separated from the PRP and sent for preparation of AS-3 RBC LR. Following filtration, the RBC unit is sealed and stored at 1-6° C for 42 days.

The PRP is centrifuged at room temperature using a “hard spin” to concentrate the platelets.

All but approximately 50 mL of plasma is expressed into the second satellite bag.

The supernatant (platelet poor) plasma is stored at temperatures of less than -20° C as Fresh Frozen Plasma, LR for up to 12 months. The plasma may also be shipped as recovered plasma for further manufacture.

The platelet units rest at room temperature for one to two hours to recover from the preparation manipulations. The platelet units are then stored on a platelet agitator in a room temperature incubator.

Additional information on Platelets, LR may be found in the Circular of Information for the Use of Human Blood and Blood Components -2004

 

Preparation of Platelet Apheresis

Platelet Apheresis, LR are collected from single donors using automated apheresis techniques, which include steps to separate leukocytes. Platelet Apheresis, LR are supplied in one 400mL container.

Additional information on Platelet Apheresis, LR, may be found in the Circular of Information for the Use of Human Blood and Blood Components - 2004 and 2005

Preparation of CPD Platelets

Below is a simple schematic of how CPD Platelets Pooled, LR are prepared from a whole blood collection using the buffy coat production method. Not shown is the procedure for leukofiltration.

Step 1		The whole blood donation is centrifuged using a “hard spin” to separate red cells from plasma. The buffy coat layer containing the platelets appears as a thick white layer above the red cells.
Step 2		Using the component separator (Compomat G4) RBCs and plasma are extracted from the buffy coat (from the bottom and top of the primary bag respectively). Red cells and plasma are forwarded for further processing.
Step 3		The buffy coats are allowed to rest for a minimum of two hours. Four buffy coats with the same ABO group and plasma from one of the same four buffy coats are grouped together for pooling.
Step 4		The buffy coats and plasma are sterile docked together in the “train” method. The platelet storage container with filter is sterile docked to the last buffy coat in the train.
Step 5		The buffy coats and plasma are pooled together using the “train” method.
Step 6		Pooled buffy coats are centrifuged using a “soft spin” to separate platelet rich plasma from the plasma containing the majority of the white blood cells.
Step 7 (picture unavailable)		Platelets (platelet rich plasma) are extracted and filtered using the Compomat G4
Step 8		CPD Platelets, Pooled, LR is similar in physical size to an apheresis platelet concentrate The platelets are placed on a platelet agitator for storage. Sampling for potential bacterial contamination is obtained at this time

Additional information on CPD Platelets, Pooled, LR may be found in the Circular of Information for the Use of Human Blood and Blood Components - 2005 

Preparation of Cryoprecipitate and Cryosupernatant Plasma

Canadian Blood Services produces two products from Fresh Frozen Plasma, LR

• Cryosupernatant, LR (Leukocytes reduced), which is also known as CSP, LR
• Cryopreciptate, LR (Leukocytes reduced), which is also known as Cryo, LR

Following the implementation of the Buffy Coat Production Method (BCPM), these components produced from CPD Frozen Plasma will also be available:

• CPD Cryosupernatant Plasma
• Cryoprecipitate

Preparation of CSP, LR and Cryo, LR

Cryoprecipitate is so named because it precipitates when frozen plasma is thawed at low temperatures. Below is a simple description of how Cryoprecipitate, LR and Cryosupernatant, LR is prepared from CP2D whole blood. Leukofiltration is not shown.

Step 1		The whole blood donation is centrifuged to separate red cells from plasma.
Step 2		Approximately 190-260 mL of donor plasma is expressed into the satellite bag. The RBC unit is sealed and stored at 1-6o C for varying shelf lives. Not shown is the AS-3 container should the RBC be made into an AS-3 RBC.
Step 3		Donor plasma is rapidly frozen within eight hours of collection to preserve Factor VIII.
Step 4		Donor plasma is thawed slowly at 1-6°C to the slush stage. Plasma is then centrifuged to separate the cryoprecipitated portion (i.e., cryoprecipitate) from the liquid portion (cryosupernatant plasma).
Step 5		All but ~10 mL of plasma is expressed into the final satellite bag. The three mL of Cryo remains in a plasma volume of about 5-15 mL. Plasma separated from the Cry is know as Cryosupernatant Plasma and can be < href="/resources/books/vein-vein/pretransfusion/storage-blood-componants">stored frozen for u to 12 month t temperatures less than minus 20OC. (While at Canadian Blood Services) Cryo i href="/resources/books/vein-vein/pretransfusion/storage-blood-componants">stored frozen for u to 12 month t 20OC or colder (whil store at CBS).   Preparat on of CPD Cryosupernatant Plasma and CPD Cryoprecipitate CPD Cryos pernatant Plasma is th newly froze roduc obtained following th production of CPD Cryo using the Buffy Coat Production Method (BCPM). Below is simple schematic of how CPD Cryoprecipitate and CPD Cyrosupernan Plasma is prepared fro CPD Whole Blood.

Buffy Coat Component Production Method

A project update from Susan Shimla, Buffy Coat Project Manager, January 2009:
The Buffy Coat production method has been successfully implemented at eleven sites, Edmonton, British Columbia & Yukon, Central Ontario, Hamilton, Calgary, Halifax, New Brunswick, Newfoundland, Ottawa, London and Winnipeg.

The last scheduled implementation occurred on November 28, 2008. All CBS sites, with the exception of Saskatchewan have converted to the new production method.

In the short term Saskatchewan will continue to produce an ongoing national supply of platelets made by the Platelet Rich Plasma method to provide a low volume platelet dose. A longer term solution, the splitting of apheresis platelets, will be investigated but is not expected to be implemented prior to March 2010.

The Buffy Coat project will officially end just prior to the end of the fiscal year with the transition of functions back into operations.

Canadian Blood Services is introducing a new method to produce platelets from whole blood collections called the Buffy Coat Component Production method. This method offers significant benefits for hospitals: a pooled, bacterially tested, ready-to-transfuse platelet concentrated with a higher yield of platelets, greater availability and a five-day shelf life.

This page summarizes the resources available on TransfusionMedicine.ca that will help you better understand this new production method. We welcome your questions and resource suggestions; please use the feedback button.

For a brief summary of changes and components, visit:

• Changes Affecting Hospitals and Patients Following the Implementation of Buffy Coat Production Method
• Amendments 1 and 2 from the 2005 Circular of Information, which lists additional information for red blood cell products manufactured by the Buffy Coat Production method.

For more general information about component preparation, visit our Component Preparation section, which has been updated to include the new products available and their storage conditions:

• Component Preparation
• Preparation of Red Blood Cells
• Preparation of Plasma Products
• Preparation of Platelets
• Preparation of Cryoprecipitate and Cryosupernatant Plasma
• Storage of Blood Components

Training Resources for Nurses and Paramedicals

For nurses and other paramedical personnel in Canadian Hospitals where the method is being introduced, we offer the following training resources:

• Blood Bag Spiking Procedures. A six-minute video showing the spiking technique for the new blood and blood product bags used in buffy coat production. Produced and hosted by the B.C. Blood Coordinating Office.
• Spiking Procedure for Use with Baxter and Macopharma Blood Bags. A presentation by Susan Shimla, Project Director, that shows the spiking technique for specific vendor blood infusion administration sets. December 2005.
• Procedure for Spiking New Buffy Coat Blood Bags. A letter sized poster with troubleshooting tips. Suitable for printing. Updated Dec 2007.
• Introducing ...... Buffy Coat Production Method. Buffy Coat Syllabus Poster. Includes benefits. August 2006.
• Buffy Coat Coming Soon to Newfoundland and Labrador Hospitals An overview article, "Do the Twist," suitable for local hospital or regional publications or newsletters. June 2008.
• Buffy Coat Project: Compatibility of transfusion sets with component bags used by the CBS. A summary of a two part study performed by the CBS designed to assess the compatibility of commonly used transfusion sets with various component bags currently used. January 2006. Important Note: Effective January 2007 all MacoPharma component bags used by CBS are of the modified port design.
• CBS Buffy Coat Blood Bag Evaluation: Technical Report. Results and recommendations from the study by the Provincial Blood Coordinating Office, B.C., investigating the introduction of new blood bags to B.C. hospitals. October 2006. Important Note: Effective january 2007 all MacoPharma component bags used by CBS are of the modified port design.
• Buffy Coat Project: Compatibility of transfer sets with component bags used by the CBS. A summary of a study performed by the CBS to assess the compatibility of commonly used transfer sets with various component bags. February 2007.
• Buffy Coat Readiness Checklist. Is your hospital ready for Buffy Coat? A sample checklist to ensure readiness for Buffy Coat products.