Storage of red blood cells affects membrane composition, microvesiculation, and in vitro quality

BackgroundDuring storage detrimental biochemical and biomechanical changes occur within a red blood cell (RBC). RBC microparticles (RMPs) produced during storage have been identified as biomarkers of RBC quality, being potentially immunogenic and inhibitory to nitric oxide regulation. Study Design and MethodsIn this study, microvesiculation and changes in the composition of the RBC membrane were investigated throughout 49 days of storage and were correlated with in vitro assays examining membrane quality. Leukoreduced RBC units produced using the buffy coat method were collected and stored at 1 to 6 degrees C and were tested weekly for hemolysis, osmotic fragility, deformability, ATP, hematologic indices, and morphology. Microvesiculation was assessed using multicolor flow cytometry. High-performance liquid chromatography and mass spectrometry were used to determine the composition and quantity of phospholipids (PLs) and cholesterol (C) on Days 2 and 43. ResultsThe assessment of RBCs throughout storage revealed significant increases in percent hemolysis, while significant decreases in ATP concentrations, and the mean corpuscular hemoglobin concentration were observed. Flow cytometry analysis revealed a significant increase in the mean number of microparticles per microliter during storage. Throughout storage, significant decreases were identified in the amount of PLs and total lipids within the RBC membrane. No significant change in the amount of C in the RBC membrane was identified. ConclusionSignificant changes to the RBC membrane occur during storage. The length of storage will influence RMP generation, osmotic fragility, hemolysis, and changes in deformability. These changes in RBC in vitro quality may contribute to transfusion reactions and negative posttransfusion outcomes.