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Exp Dermatol. 2014 Feb;23(2):125-9. doi: 10.1111/exd.12322.

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

Author information

1
Department of Dermatology, Cutaneous Biology Research Center, Massachusetts General Hospital, Harvard Medical School, Boston, MA, USA; Department of Dermatology, Cutaneous Biology Research Institute, Yonsei University College of Medicine, Seoul, Korea.

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.

KEYWORDS:

co-culture; high-content screening; keratinocyte; melanocyte; microphthalmia transcription factor; pigmentation

PMID:
24438532
PMCID:
PMC3977999
DOI:
10.1111/exd.12322
[Indexed for MEDLINE]
Free PMC Article

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