Surface area and volume changes during maturation of reticulocytes in the circulation of the baboon

Changes in the surface area and volume of reticulocytes were measured in vivo during late stage maturation. Baboons were treated with erythropoietin to produce mild reticulocytosis. Reticulocyte-rich cohorts of cells were obtained from whole blood by density gradient centrifugation. The cohorts were labeled with biotin, reinfused into the animal, and recovered from whole blood samples by panning on avidin supports. Changes in the surface area, volume, and membrane deformability were measured using micropipettes during the 2 to 6 weeks subsequent to reinfusion. For the entire cohort, the membrane area decreased by 10% to 15% and the cell volume decreased by approximately 8.5%, mostly within 24 hours after reinfusion. Estimates of the cellular dimensions of the reticulocyte subpopulation within this cohort indicated larger reductions in the mean cell area (12% to 30%) and mean cell volume (approximately 15%) of the reticulocytes themselves. Two weeks after reinfusion, the distribution of cell size for the cohort was indistinguishable from that of whole blood. There was evidence of slightly elevated membrane shear rigidity in some reticulocytes before reinfusion, but this slight increase disappeared within 24 hours after reinfusion. These are the first direct measurements of changes in the membrane physical properties of an identifiable cohort of reticulocytes as they mature in vivo.