Analysis of density-fractionated rat red blood cells of different ages by partitioning in two-polymer aqueous phase systems.
Rats were injected with radioactive iron and bled at different times thereafter. This gave rise to cell populations in which the bulk of labeled cells corresponded in age to the time elapsed between injection and bleeding. Such cell populations were centrifuged and the 7-10% least dense and the 7-10% most dense cells subjected to counter-current distribution in a charge-sensitive two-polymer aqueous phase system which fractionates cells on the basis of surface properties. While it is known that there is a tendency for cell density to increase with cell age and that rat red cells of different ages have characteristic partition ratios, we were able, by applying partitioning analysis to density-separated cells, to gather data not obtainable by use of either method alone. These include: (1) the demonstration of two very young cell populations having different surface properties in the most dense fraction; (2) the finding that in the least dense layer labeled mature erythrocytes (at least those labeled 18 days and beyond) and unlabeled red cells have overlapping distribution curves which indicate that the label in the top fraction can represent young or early middle-aged but never old cells; and (3) the presence, as judged by surface properties, of some middle-aged red cells, in addition to old cells, in the most dense cell layer.