The Separation and Characterization of Young and Old Erythrocytes

Abstract

Results of an investigation by Huisman and Meyering (1960) of Hb-Ar in young and old cells of two adults and three infants using eM-cellulose chromatography showed that the old red blood cells contained considerably more Hb-Ar than younger red cells. Similar studies by Huisman and Horton (1964) on a more limited scale and with the use of Arnberlite IRe-50 chromatography failed to demonstrate differences in the quantities of the Hb-Ar components in hernolysates of young and old cells. Because of this discrepancy it was considered of importance to extend these observations by studying the young and old red blood cells of a relatively large number of normal healthy adult individuals. The Amberlite IRe-50 chromatographic procedure was selected for this purpose because this technique was found to have the advantage over eM-cellulose chromatography of a better differentiation of the Hb-A1a, Hb-A1b, and Hb-Arc components, while the complex of Hb with -SG groups could be demonstrated as a separate component (Hb-Ard) with its own·specific chromatographic properties. In order to assure a definite fractionation of the adult red blood cells into layers of cells with different cell-age--a-new procedure was developed, which moreover, allowed us sufficient quantities of (intact) ·cell fractions for additional characterization.