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Growth characteristics of human epidermal melanocytes in pure culture with special reference to genetic differences
Author(s) -
Hirobe Tomohisa,
Flynn Evelyn,
Szabo George,
Vrabel Michael,
Garcia Raul I.
Publication year - 1988
Publication title -
journal of cellular physiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.529
H-Index - 174
eISSN - 1097-4652
pISSN - 0021-9541
DOI - 10.1002/jcp.1041350213
Subject(s) - melanocyte , melanosome , melanin , in vitro , biology , cell culture , epidermal growth factor , microbiology and biotechnology , fetal bovine serum , melanocyte stimulating hormone , cell growth , endocrinology , hormone , biochemistry , melanoma , genetics
Human melanocyte cultures were established using disaggregated epidermal cell suspensions derived from foreskins and plated onto culture dishes in medium containing 2% fetal bovine serum, growth factors, hormones, and melanocyte growth factor (MGF) extracted from bovine hypothalamus (Wilkins et al., J. Cell. Physiol., 122 : 350, 1985). After 2 days in culture the cells were transferred to serum‐free medium to eliminate keratinocyte and fibroblast growth. Melanocytes grew preferentially and pure melanocyte populations could be harvested after 12–16 days in vitro. Melanocytes were later subcultured in the presence of 1% FBS. Pure melanocyte cultures were characterized by light and electron microscopic criteria, as well as by cytochemical demonstration of the melanocytes specific enzyme, tyrosinase. At the ultrastructural level, cultured melanocytes derived from black (negroid) neonatal skin (B‐M) had numerous mature rodshaped stage IV melanosomes, while white (caucasoid) skin‐derived melanocytes (W‐M) in culture contained no mature melanosomes. Growth rate, cell yield, and in vitro lifespan for B‐M were more than twice that for W‐M in pure melanocyte cultures in the presence of MGF. Our results suggest that MGF‐dependent growth of B‐M differs from that of W‐M.