Influence of temperature and method of centrifugation on the separation of erythrocytes.

Abstract

Abstract Previous studies have demonstrated that erythrocytes can be separated on the basis of specific gravity by centrifugation in various media. Horizontal centrifugation with swinging bucket rotors at 0 ° to 5 °C. was invariably used in these earlier studies. The use of discontinuous gradients with centrifugation has been more effective in separating cells by specific gravity, but is limited in the volume of cells that can be separated and is time consuming to prepare. The method described in this study utilizes the internal circulation in a tube centrifuged in an angle-head rotor at 30 ° C. at which the erythrocyte is more deformable as compared to 0 to 5 °C. Angle-head centrifugation at 30 °C. ± 2 ° for 1 hour resulted in separation of normal cells to the extent that the bottom 5 per cent of the cells had a mean corpuscular hemoglobin concentration (MCHC) of 124 per cent that of the top 5 per cent of the cells. In sickle cell anemia the MCHC of the bottom cells was 160 per cent that of the top cells. This improved method for separating erythrocytes on the basis of specific gravity, which has previously been shown to be related to cell age, will provide a means for more definitive observations of cell aging.