Cryopreservation of red blood cell units with a modified method of glycerolization and deglycerolization with the ACP 215 device complies with American and European requirements

Current red blood cell (RBC) glycerolization with the ACP 215 device is followed by volume reduction of the glycerolized RBCs before freezing. We investigated a modified method of glycerolization and deglycerolization which eliminates the final centrifugation step that reduces glycerolized RBC supernatant. A total of 37 RBC units collected from healthy volunteers were analyzed. After removal of the supernatant, RBCs were glycerolized using the high glycerol method and stored at –80°C. After deglycerolization, RBCs were preserved with either SAG-M or AS3 and stored for at least 10 or 14 days, respectively. Quality of stored RBCs was assessed by measuring osmolarity, blood cell counts, free hemoglobin, adenosine triphosphate (ATP), hemolysis, and glucose. The overall RBC mass recovery after deglycerolization was 86 ± 7.6 percent, and the osmolarity was 336 ± 23 mOsml/kg H₂O. The hemolysis for stored components at the end of their shelf life was 0.21 ± 0.08 percent for AS3-preserved RBCs and 0.25 ± 0.08 percent for RBCs preserved with SAG-M. On expiration, 32 percent of initial ATP values were measured in AS3-preserved RBCs vs. 62 percent in SAG-M–preserved RBCs. This modified method of glycerolization and deglycerolization meets the quality requirements of the European Council and the AABB standards. The prolonged storage of thawed RBCs enables optimized transfusion management for patients with rare blood groups. Immunohematology2012;28:67–73.