Susceptibility to hyperosmotic stress-induced phosphatidylserine exposure increases during red blood cell storage

During storage of red blood cell (RBCs) before transfusion, RBCs undergo a series of structural and functional changes that include the exposure of phosphatidylserine (PS), a potent removal signal. It was postulated that, during blood bank storage, the susceptibility to stress-induced PS exposure increases, thereby rendering a considerable fraction of the RBCs susceptible to rapid removal after transfusion.

RBCs were processed and stored following standard Dutch blood bank procedures. Samples were taken every week for up to 6 weeks and exposed to various stress conditions, such as hyperosmotic shock and energy depletion. The effect of these treatments on PS exposure was measured by flow cytometric analysis of annexin V binding. The same analyses were performed on RBCs that had been separated according to density using discontinuous Percoll gradients.

During storage under blood bank conditions, RBCs become increasingly susceptible to loss of phospholipid asymmetry induced by hyperosmotic shock and energy depletion. Especially the RBCs of higher densities, that have a smaller volume and an increased HbA1c content as is typical of aged RBCs, become increasingly susceptible with storage time.

During storage, RBCs develop an increased susceptibility to stress-induced loss of phospholipid asymmetry that is especially associated with an aging phenotype. This increased susceptibility may be responsible for the rapid disappearance of a considerable fraction of the RBCs during the first 24 hours after transfusion.