Comparison of the human melanocyte culture in phorbol 12-myristate 13-acetate-contained medium and physiologic mitogens-contained medium / 대한피부과학회지

BACKGROUND: The growth of cells is closely related to components in a culture medium. There are many reports about cellular characteristics of melanocytes grown in a PMA-contained medium. However, only a few reports have been studied by using a physiologic mitogens-contained medium. To understand melanocyte in vivo, it is necessary to know the cellular biology of melanocytes grown in a physiologic mitogens-contained medium. OBJECTIVE: To investigate any differences between melanocytes grown in phorbol 12-myristate 13-acetate(PMA)-contained medium and in physiologic mitogens-contained medium. METHOD: We examined morphology, number and melanin contents of cultured human melanocytes grown in a PMA-contained medium and physiologic mitogens-, such as bFGF, ET-1 and a a-MSH contained medium. Result : The results are summarized as follows : 1. There were no significant morphologic differences between cells in PMA-contained medium and in physiologic mitogens-contained medium. 2. The number of melanocytes were significantly more numerous in PMA-contained medium on the 2nd day (p<0.05), but significantly less numerous in the same medium on the 6th day (p<0.05). So, the proliferation rate of melanocytes in PMA-contained medium became lower than in physiologic mitogens-contained medium as time went by. 3. Melanocytes grown in PMA-contained medium had significantly increased melanin contents regardless of the time (p<0.05). Conclusion : The proliferation of melanocytes was better in physiologic mitogens-contained medium, the melanization was higher in melanocytes of PMA-contained medium.

Effect of mitogenic factors extracted from fetal lung fibroblasts on the in vitro growth of melanocytes obtained from normal and vitiligo subjects.

The effects of growth factors extracted from a newly established fetal lung fibroblast cell line (PMR-GF) on the melanocytes cultured from the perilesional and depigmented skins of vitiligo subjects and from normal healthy donors have been investigated. Melanocytes from normal subjects grown in the presence of 10 ng/ml of 12-0-tetradecanoyl phorbol 13-acetate and 10–11 Μ cholera toxin grew exponentially immediately after seeding the epidermal cell suspensions. Exogenous addition of PMR-GF to these cells enhanced their growth rates. The perilesional skin melanocytes of vitiligo subjects in most cases did not manifest any growth when cultured in the presence of 12-0-tetradecanoyl phorbol 13-acetate and cholera toxin. PMR-GF induced a brief burst of growth in these cells after a lag of 15 days. Vitiligo lesions gave rise to a few unpigmented dendritic cells that did not manifest any growth in the presence or absence of PMR-GF. Morphologically the perilesional skin melanocytes of most vitiligo subjects, when cultured in 12-0-tetradecanoyl phorbol 13-acetate and cholera toxin, appeared to be larger and hyper-melanotic as compared to those of normal individuals. In the presence of PMR-GF these melanocytes appeared to be normal in size and less hypermelanotic. Our results indicate that the melanocytes from vitiligo subjects are defective and thus the basic defect in vitiligo could be with the melanocytes themselves.