IMPROVED RED-BLOOD-CELL PRESERVATION CORRELATES WITH DECREASED LOSS OF BAND-3, BAND-4.1, ACETYLCHOLINESTRASE, AND LIPIDS IN MICROVESICLES

In earlier studies we have shown that a final concentration of 0.69% g lycerol in blood mixed with an experimental additive solution, EAS 25, improves the in vitro quality and in vivo survival of red blood cells (RBCs), the objective of this study was to determine if the better pr eservation of RBCs in EAS 25 is correlated with the improved maintenan ce of membrane lipids and proteins and decreased vesiculation, Split u nits of RBCs were stored in Adsol or EAS 25 (mmol/L: adenine 2/2, dext rose 122/110, mannitol 42/55, glycerol 0/150, NaCl 154/50). After 12 w eeks storage, RBC and microvesicle membranes were analyzed for cholest erol, phospholipid, diphenyl hexatriene fluorescence anisotropy, and a cetylcholinesterase (AchE) activity, Bands 3 and 4.1 were identified i n the microvesicle membranes by immunoblotting. The RBC membrane chole sterol, phospholipids, and AchE remained higher in EAS 25 than in Adso l (P<.001). Vesicle membrane lipids and AchE in EAS 25 were significan tly less than in Adsol (P<.001). The fluidity of stored cells in both the solutions was greater than the prestorage samples. Immunoblotting analyses showed that bands 3 and 4.1 were greatly reduced in the micro vesicle membranes shed by the RBCs stored in EAS 25 compared with thos e formed in Adsol. (C) 1996 by The American Society of Hematology.