This is a Standard Operating Procedure (SOP) to glycerolize and deglycerolize human red blood cells using the Haemonetics ACP215 automated, closed system. The red blood cells can be collected into any FDA-approved anticoagulant or anticoagulant-preservative system. Prior to freezing the whole blood or additive-stored red blood cells are concentrated to prepare a red blood cell concentrate with a hematocrit value of approximately 75 V%. If the blood was not collected into the 800 ml PVC plastic bag collection system, the red blood cells are transferred into a standard 1000 ml polyvinyl chloride plastic bag using a sterile docking procedure. The red blood cells are glycerolized using the ACP215 and the glycerolized red blood cells are concentrated and the supernatant glycerol removed prior to freezing. The glycerolized red cells are washed in the Haemonetics ACP215 and resuspended in AS3 solution for post-wash storage.

The ACP215 system provides a functionally closed system for both the glycerolization and deglycerolization procedure resulting in a reduced potential for bacterial contamination. This, in addition to the resuspension of the deglycerolized red blood cells in AS-3 solution, allows for storage of the red blood cells at 4 C for up to 14 days following washing.