Washing Packed Red Blood Cells During Storage Decreases Storage Lesion Formation
*Kasiemobi Pulliam, *Bernadin Joseph, *Rosalie A Veile, Amy T Makley, *Charles C Caldwell, *Alex B Lentsch, Michael D Goodman, Timothy A Pritts
University of Cincinnati, Department of Surgery, Cincinnati, OH
OBJECTIVES: In traumatic hemorrhage, transfusion of blood products is the preferred method of resuscitation. Unfortunately, massive transfusion with older packed red blood cells (pRBCs) has been associated with increased morbidity and mortality after trauma. Packed red blood cell storage is associated with accumulation of biochemical and structural changes to the erythrocyte, known as the red blood cell storage lesion. We have previously shown that washing pRBCs at the time of transfusion reduces inflammatory mediators, but the effect of washing on red blood cell storage lesion formation is unknown. We hypothesized that washing pRBCs during storage will reduce the development of the red blood cell storage lesion.
METHODS: Blood from 8-10 week old C57BL/6 male mice donors was obtained and stored as pRBCs under standard storage conditions for 14 days. A subset of standard pRBCs were washed with 1X PBS on storage day 7 and resuspended in AS-3 for an additional 7 days as washed pRBCs. At this time, standard and washed pRBCs were analyzed for microvesicle formation, band 3 erythrocyte membrane integrity protein (Band-3), phosphatidylserine expression (PS), cell viability (calcein), cell-free hemoglobin accumulation, and susceptibility to osmotic stress (EC50).
RESULTS: There was less microvesicle accumulation (6406.6 + 402.8 vs. 2915.0 + 647.1 events/μL) in the washed pRBCs group. The washed pRBCs demonstrated greater Band-3 expression (37.8 + 0.65 vs. 92.06 + 0.58 % gated) and less phosphatidylserine expression (52.9 + 1.3 vs. 58.7 + 0.4 % gated). There was less cell free hemoglobin accumulation (49.4 + 3.4 vs. 41.6 + 2.9 g/dL). There were no differences in RBC viability (125018 + 3384.7 vs. 131673 + 1160 MFI) nor susceptibility to osmotic stress (EC50 0.83 + 0.04 vs. 0.90 + 0.01; all results p<0.05 vs standard).
CONCLUSION: Washing pRBCs during storage reduces the accumulation of the red blood cell storage lesion. This strategy may lessen the sequelae associated with transfusion of older pRBCs.
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