A melanocyte-keratinocyte coculture model to assess regulators of pigmentation in vitro.
Many melanocyte or skin equivalent models have been used to evaluate the potential efficacy of melanogenic compounds to regulate pigmentation, but there has been great variation in results, partially stemming from the use of different cell lines and diverse conditions for the melanogenic assays. In an earlier report, we optimized a microtiter format assay system to screen potential bioactive compounds using immortalized melan-a melanocytes. That assay system, termed the STOPR protocol, allowed effects on melanocyte proliferation and differentiation to be assessed in a highly sensitive, reproducible, and cost-effective manner. However, in the skin and hair, melanocytes interact with keratinocytes, fibroblasts, and other cell types, and testing of putative bioactive compounds on melanocytes alone in culture does not allow one to observe the interactions with those other cell types, such as would occur in vivo. Therefore, we developed a melanocyte-keratinocyte coculture protocol that allows testing of compounds for potential effects on pigmentation in a more physiologically relevant context. It is a sensitive, reproducible, and reliable model for testing melanogenic regulators, and we have standardized it with known melanogenic inhibitors (hydroquinone, arbutin, kojic acid, and niacinamide) and stimulators (alpha-melanocyte-stimulating hormone, 8-methoxypsoralen, and 3,4-dihydroxyphenylalanine). This coculture system allows for large-scale screening of candidate compounds in conjunction with the STOPR protocol and provides a more physiologically relevant system to study melanocyte-keratinocyte interactions and to elucidate the regulatory mechanisms of melanogenic compounds.
Lei
Tie Chi
TC
Pigment Cell Biology Section, Laboratory of Cell Biology, Bethesda, Maryland 20892, USA.
Virador
Victoria M
VM
Vieira
Wilfred D
WD
Hearing
Vincent J
VJ
D000818:Animals; D001104:Arbutin / Q000494:pharmacology; D018920:Coculture Techniques / Q000379:methods; D004295:Dihydroxyphenylalanine / Q000378:metabolism; D006873:Hydroquinones / Q000494:pharmacology; D015603:Keratinocytes / Q000187:drug effects / Q000378:metabolism*; D009074:Melanocyte-Stimulating Hormones / Q000378:metabolism; D008544:Melanocytes / Q000187:drug effects / Q000378:metabolism*; D051379:Mice; D008954:Models, Biological; D014442:Monophenol Monooxygenase / Q000037:antagonists & inhibitors; D009536:Niacinamide / Q000494:pharmacology; D010858:Pigmentation / Q000502:physiology*; D011753:Pyrones / Q000494:pharmacology
D006873:Hydroquinones; D011753:Pyrones; D009536:Niacinamide; D004295:Dihydroxyphenylalanine; C011890:kojic acid; D009074:Melanocyte-Stimulating Hormones; D001104:Arbutin; D014442:Monophenol Monooxygenase; C031927:hydroquinone