Altered processing of thawed red cells to improve the in vitro quality during postthaw storage at 4 degrees C.

BACKGROUND: The use of a functionally closed system (ACP215, Haemonetics) for the glycerolization and deglycerolization of red blood cell (RBC) units allows for prolonged postthaw storage. In this study, the postthaw quality of previously frozen, deglycerolized RBCs resuspended in saline-adenine-glucose-mannitol (SAGM) or additive solution AS-3 was investigated. STUDY DESIGN AND METHODS: Leukoreduced RBC units were frozen with 40 percent glycerol and stored at -80 degrees C for at least 14 days. The thawed units were deglycerolized with the ACP215, resuspended in SAGM or AS-3, and stored at 2 to 6 degrees C for up to 21 days. RESULTS: The mean +/- standard deviation in vitro freeze-thaw-wash recovery was 81 +/- 5 percent. During storage, hemolysis of deglycerolized cells remained below 0.8 percent for 2 days in SAGM and for 14 days in AS-3. This difference was explained by the protective effect of citrate, which is present in AS-3. Cells stored in AS-3 showed a lower glycolytic activity and a faster decline in adenosine 5'-triphosphate (ATP) than cells in SAGM. Increasing the internal pH of cells before storage in AS-3 by use of phosphate-buffered saline (PBS) in the deglycerolization procedure resulted in elevated lactate production and better maintenance of intracellular ATP content. After 3 weeks of storage, the ATP content of PBS-washed cells amounted to 2.5 +/- 0.5 micromol per g of hemoglobin (Hb), whereas for saline/glucose-washed cells this value was decreased to 1.0 +/- 0.3 micromol per g of Hb. CONCLUSIONS: Leukoreduced, deglycerolized RBCs can be stored for 48 hours in SAGM. Improved ATP levels during refrigerated storage can be observed with thawed cells, resuspended in AS-3, when PBS is used as a washing solution.

Stability after thawing of RBCs frozen with the high- and low-glycerol method.

BACKGROUND: RBCs can be frozen with either the high-glycerol method (HGM) or the low-glycerol method (LGM). To date, the use of frozen RBCs is hampered by a 24-hour outdating period after thawing. A closed washing system (ACP 215) may solve this problem. STUDY DESIGN AND METHODS: We compared the effects of high- (40%) and low-glycerol (19%) concentration, with and without freezing (at -80 degrees C for HGM, -196 degrees C for LGM) on the in vitro quality of RBCs after deglycerolization with the closed washing system and during storage at 4 degrees C in SAGM after thawing. RESULTS: Glycerol treatment by itself induced hemolysis during processing, which was more pronounced in HGM cells. The freeze-thaw-wash process decreased the stability of RBCs, particularly in LGM cells during storage after thawing. In contrast to LGM cells, in HGM cells no additional effect of freeze or thaw on stability of washed cells was seen during the first week of storage after thawing. Changes in osmotic resistance and cellular metabolism could not explain the observed differences in RBC stability. CONCLUSION: The closed washing system is able to process both high- and low-glycerol-treated RBCs. Stability after washing during cold storage in SAGM, as measured by hemolysis, is better for HGM cells as compared to LGM cells.