A new medium for primary cultures of adult murine epidermal cells: application to experimental carcinogenesis

Abstract

A culture medium has been formulated to support the long-term proliferation and differentiation of normal murine epidermal cells in primary cultures. This formulation is based upon MCDB-151 modified by the addition of 1.2 mM Ca2+, 1% fetal bovine serum (FBS) or 0.1% purified bovine serum albumin (BSA), trace elements, and growth supplements. For 4-12 weeks, the cultures demonstrate desmosomes, keratin fibers, basal proliferation, and stratification characteristic of keratinocytes. The addition of 12-O-tetradecanoylphorbol-13-acetate (TPA) resulted in approximately 70% cellular detachment, but with continued treatment, the remaining cells proliferated to confluence and stratified. This culture medium has application to a number of studies related to skin carcinogenesis in the murine model system that have not previously been possible.