Studies on thymocyte subpopulations in guinea pigs: in vivo differentiation of bromodeoxyuridine labelled cells, with special reference to rosette-forming ability

Cycling thymocytes were labelled by an intracardial injection of bromodeoxyuridine (BrdUrd) in a total of 32 guinea pigs and the incorporation into DNA studied in subpopulations of cells defined by buoyant density and rosette-forming ability. The labelling pattern was compared at different times after injection (0.5 h to 120 h). A marked shift of labelled cells from large, low density cells (population 1a) to small, high density cells (population 2) was observed. During the first 48 h, the ratio between labelling indices of cell populations 1a and 2 decreased from 10 to 0.5. The number of labelled cells forming rosettes with rabbit erythrocytes (RFC+) increased while the number of labelled non-rosetting cells (RFC-) decreased from 0.5 h to 48 h, probably representing transformation of RFC- to RFC+. Then, after a decreased labelling in all cell populations at 72 h, an increase in both RFC+ and RFC- populations occurred at 96 h. The labelling in RFC- cells at 96 h was nearly as high as immediately after labelling. This second labelling of RFC- cells could represent immigration of precursor cells, a wave of proliferation in initially labelled precursors, and/or the formation of mature cells from the initially labelled precursors. The results indicate that a great majority of proliferating cells differentiate into small, high density cells within 48 h and that rosetting ability is acquired in many cells during this period. A model of intrathymic differentiation which fits the observations is presented.