Organotypic Skin Model Containing Melanocytes

Many highly differentiated in vitro epidermal and dermal/epidermal skin models have beendeveloped which are useful for several reasons: 1) transdermal drug delivery systems, 2) skintransplantation, and 3) dermal irritancy testing. For the most part, these models incorporate epidermalkeratinocytes and dermal fibroblasts, but do not include melanocytes, the pigment cells of theintegument which provide protection from potentially harmful ultraviolet (UV) solar irradiation. Inaddition, melanocytes are believed to participate in the inflammatory response of skin and are obviouslyrequired to allow the study of skin pigmentation disorders, UV induced skin damage, etc. Utilizing aserum free, highly differentiated keratinocyte based epidermal model previously developed as a startingpoint, this research will attempt to develop a similar serum free culture system which allows theincorporation of melanocytes. A serum free medium will be developed using additives known tostimulate the proliferation and maintenance of melanocytes in monolayer culture. Three dimensional,highly differentiated epidermal constructs will be cultured and characterized using light and electronmicroscopy, a melanocyte biochemical marker, and melanin synthesis. Finally, an initial investigationof the model's response to stimuli known to affect melanocytes in vivo such s UV will be made. Thesestudies will lay the groundwork for Phase II research which will include an in-depth analysis of the effectof UV on the epidermis, photoaging of skin, and other melanocyte related skin phenomena.