Reconstituted 3-dimensional human skin of various ethnic origins as an in vitro model for studies of pigmentation
Section snippets
Chemicals and solvents used
MSH, DOPA, hydroquinone, arbutin, kojic acid, niacinamide, phorbol 12-myristate 13-acetate (PMA), and β-mercaptoethanol were from Sigma Chemical (St. Louis, MO). l-[U-14C]tyrosine was from Amersham Biosciences (Piscataway, NJ). All compounds were dissolved in water and were protected from light at −20 °C until used, except PMA, which was dissolved in DMSO.
Culture and treatment of pigmented reconstituted human epidermis
MelanoDerm (MatTek Corp., Ashland, MA) is a viable reconstituted 3-dimensional human epidermis containing melanocytes and keratinocytes that
General considerations
We have previously established streamlined protocols (the STOPR and the co-culture models) that use immortalized mouse melanocytes with or without keratinocytes in monolayer culture to measure effects on pigmentation [18], [19]. These 2-dimensional models do not completely reflect the physiological effects of bioactive compounds, because melanocyte or melanocyte–keratinocyte interactions are limited spatially in 2-dimensional monolayers, rather than the 3-dimensions that occur in the skin. In
Conclusions
Numerous pigmented melanoma or melanocyte culture models have been used (some with co-cultures of keratinocytes) for evaluating the efficacy of melanogenic compounds [18], [19], [35], [36], but each has only limited value when compared with physiologically relevant, pigmented 3-dimensional skin equivalents. The 3-dimensional skin models have the advantage that they show an accurate histological structure and allow one to assess not only pigment synthesis, but pigment translocation and melanin
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Present address: Department of Dermatology, Gyeongsang National University Hospital, 90 Chilam-Dong, Chinju, Gyeongnam 660-702, South Korea.