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High-throughput, high-content screening for novel pigmentation regulators using a keratinocyte/melanocyte co-culture system.


ABSTRACT: Skin pigmentation is a complex process including melanogenesis within melanocytes and melanin transfer to the keratinocytes. To develop a comprehensive screening method for novel pigmentation regulators, we used immortalized melanocytes and keratinocytes in co-culture to screen large numbers of compounds. High-throughput screening plates were subjected to digital automated microscopy to quantify the pigmentation via brightfield microscopy. Compounds with pigment suppression were secondarily tested for their effects on expression of microphthalmia transcription factor (MITF) and several pigment regulatory genes, and further validated in terms of non-toxicity to keratinocytes/melanocytes and dose-dependent activity. The results demonstrate a high-throughput, high-content screening approach, which is applicable to the analysis of large chemical libraries using a co-culture system. We identified candidate pigmentation inhibitors from 4000 screened compounds including zoxazolamine, 3-methoxycatechol and alpha-mangostin, which were also shown to modulate expression of MITF and several key pigmentation factors and are worthy of further evaluation for potential translation to clinical use.

SUBMITTER: Lee JH 

PROVIDER: S-EPMC3977999 | biostudies-literature | 2014 Feb

REPOSITORIES: biostudies-literature

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High-throughput, high-content screening for novel pigmentation regulators using a keratinocyte/melanocyte co-culture system.

Lee Ju Hee JH   Chen Hongxiang H   Kolev Vihren V   Aull Katherine H KH   Jung Inhee I   Wang Jun J   Miyamoto Shoko S   Hosoi Junichi J   Mandinova Anna A   Fisher David E DE  

Experimental dermatology 20140201 2


Skin pigmentation is a complex process including melanogenesis within melanocytes and melanin transfer to the keratinocytes. To develop a comprehensive screening method for novel pigmentation regulators, we used immortalized melanocytes and keratinocytes in co-culture to screen large numbers of compounds. High-throughput screening plates were subjected to digital automated microscopy to quantify the pigmentation via brightfield microscopy. Compounds with pigment suppression were secondarily test  ...[more]

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