Isolation and Culture of Epidermal Melanocytes.

Melanocytes which represent around 5% of epidermal cells are located in the basal layer. To culture melanocytes we used trypsin digestion instead of dispase to obtain a cell suspension containing only basal keratinocytes and melanocytes. Melanocytes are cells which need a great attention. Indeed they dedifferentiate easily in culture as soon as they are in pure culture. Factors secreted by contaminating keratinocytes allow melanocytes to stay dendritic but by regulating their number avoid their growth. In order to age, phototype and other individual dependent factors regulate the behavior of melanocytes in vitro. Thus, microscopic examination of melanocytes has to be performed each day to adapt conditions of culture to each primary cell culture. This is the secret to have a nice melanocyte culture.