Investigations on normal human adult epidermal melanoytes

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For obtaining pure melanocyte cultures from human adult epidermis a novel culture technique using a cholera toxin and PMA-free medium (Mel-mix) was described by our group. In Mel-mix medium the cultured melanocytes become bipolar, unpigmented and highly proliferative. Further characterization of the cultured melanocytes revealed the disappearance of c-Kit and TRP-1 and induction of nestin expression, indicating that melanocytes dedifferentiated in this in vitro culture. Cholera toxin and PMA were able to induce c-Kit and TRP-1 protein expressions in the cells, reversing dedifferentiation. TRP-1 mRNA expression was induced in dedifferentiated melanocytes by UV-B irradiated keratinocyte supernatants, however direct UV-B irradiation of the cells resulted in further decrease of TRP-1 mRNA expression. Expressions of EGFR mRNA and protein and mitogenic effect of EGF on Mel-mix cultured melanocytes were proved. EGFR protein expression was more intensive in dedifferentiated melanocytes. These dedifferentiated, easily accessible cultured melanocytes provide a good model for studying melanocyte differentiation and possibly transdifferentiation. Because melanocytes in Mel-mix medium can be cultured with human serum as the only supplement, this culture system is also suitable for autologous cell transplantation.

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