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Altered processing of thawed red cells to improve the in vitro quality during postthaw storage at 4°C
Author(s) -
Lagerberg Johan W.M.,
Truijensde Lange Rosa,
De Korte Dirk,
Verhoeven Arthur J.
Publication year - 2007
Publication title -
transfusion
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.045
H-Index - 132
eISSN - 1537-2995
pISSN - 0041-1132
DOI - 10.1111/j.1537-2995.2007.01453.x
Subject(s) - cryopreservation , andrology , hemolysis , mannitol , saline , adenosine triphosphate , cold storage , glycerol , chemistry , biology , biochemistry , immunology , embryo , medicine , endocrinology , horticulture , microbiology and biotechnology
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°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°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 µmol per g of hemoglobin (Hb), whereas for saline/glucose‐washed cells this value was decreased to 1.0 ± 0.3 µmol 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.