Isolation of enriched human melanocyte cultures from fetal, newborn and adult skin

Abstract

Since melanocytes replicate slowly compared to other skin cells, it has been common practice to isolate pure melanocytes early, prior to overgrowth by fibroblasts and keratinocytes. Contact with other skin cells in culture such as keratinocytes and fibroblasts, however, can enhance melanocyte viability and promote melanocyte replication in the presence of mitogens. We routinely observed that melanocytes appeared earlier and were cultured more efficiently using primary cultures of ‘mixed’ skin cells in mitogen-containing growth medium. A method incorporating dermo-epidermal separation with thermolysin prior to epidermal digestion with trypsin proved most reliable in isolating melanocytes in mixed culture from neonatal skin samples. Pure melanocyte cultures were subsequently obtained from the primary co-culture by applying traditional separation techniques with minor modifications.