Subpopulations of primary adult murine epidermal basal cells sedimented on density gradients

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Abstract
Epidermal cells were harvested from the dorsal skin of adult mice by trypsinization and were sedimented through continuous density gradients of Percoll, formulated to separate basal cells of different buoyant density. Five fractions from the gradients were characterized with regard to the number of cells present, their viability and morphology and their basal origin. Suprabasal keratinocytes remained primarily at the top of the gradient; basal keratinocytes sedimented throughout. With increasing density, a relative enrichment was observed: (i) for [3H]-thymidine and [3H]-benzo[alpha]pyrene label-retaining (slowly cycling) keratinocytes; (ii) for keratinocytes that could proliferate in vitro in the continuous presence of 0.1 micrograms ml-1 of 12-O-tetradecanoylphorbol-13-acetate; (iii) for cells from untreated as well as initiated epidermis able to proliferate under conditions where calcium induces terminal differentiation; and (iv) for primary in vitro clonogenic keratinocytes from normal epidermis. The relative enrichment for epidermal basal cells having characteristics thought to be associated with immaturity and with the initiation and promotion of skin carcinogenesis suggests that density gradient sedimentation could be used in conjunction with other methods for the eventual purification of epidermal progenitors.