Subpopulations of Slowly Cycling Cells In S and G2 Phase In Mouse Epidermis

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Abstract
Evidence has been presented supporting the existence of heterogeneity in cell‐cycle progression in mouse epidermis, the present study was undertaken to characterize this heterogeneity in more detail. Hairless mice were continuously labelled with tritiated thymidine every 4 hr for 4 days. Basal cell suspensions were prepared from slices of mouse skin at intervals during the experiment and subjected to DNA flow cytometry. Cell‐cycle analysis was combined with sorting of cells from windows in G1, S and G2, phase, and the proportion of labelled cells within each window was determined in autoradiographs. Reanalysis and resorting to control the purity of sorted fractions were performed.Computer simulations of the data were made using a mathematical model assuming different S and G2 phase characteristics. A good fit to the data was only obtained when heterogeneity in mouse epidermal cell‐cycle progression was assumed, indicating the existence of slowly traversing, distinct subpopulations of cells in G2 and S phase. These cells are assumed to contribute to about 40% of all cells in S phase and to about 70% of all in G2 phase. the estimated residence times in the resting states were 38 and 32 hr in S and G2 phase, respectively. Two‐parameter sorting based on DNA and light scatter indicated that slowly cycling cells were larger than the average. There is no evidence of significant subpopulations of permanently non‐proliferating keratinocytes in any of the cell‐cycle phases.