title: Granulocyte-macrophage colony-stimulating factor is a keratinocyte-derived factor involved in regulating the proliferation and differentiation of neonatal mouse epidermal melanocytes in culture.
abstract: Mouse epidermal melanoblasts and melanocytes preferentially proliferated from disaggregated epidermal cell suspensions derived from newborn mouse skin in a serum-free melanocyte-proliferation medium (MDMD) and a melanoblast-proliferation medium (MDMDF) supplemented with dibutyryl adenosine 3':5'-cyclic monophosphate (DBcAMP) and/or basic fibroblast growth factor (bFGF). Pure cultured primary melanoblasts and melanocytes were further cultured with MDMD/MDMDF supplemented with granulocyte-macrophage colony-stimulating factor (GMCSF) from 14 days (keratinocyte depletion). GMCSF stimulated the number of melanoblasts/melanocytes as well as the percentage of differentiated melanocytes in keratinocyte-depleted cultures. Flow cytometry analysis showed that melanoblasts and melanocytes in the S and G(2)/M phases of the cell cycle were increased by the treatment with GMCSF. Moreover, anti-GMCSF antibody added to MDMD/MDMDF from the initiation of the primary culture (in the presence of keratinocytes) inhibited the proliferation of melanoblasts/melanocytes as well as the differentiation of melanocytes. Enzyme-linked immunosorbent assay of culture media revealed that GMCSF was secreted from keratinocytes, but not from melanocytes. These results suggest that GMCSF is one of the keratinocyte-derived factors involved in regulating the proliferation and differentiation of neonatal mouse epidermal melanoblasts/melanocytes in culture in cooperation with cAMP elevator and bFGF.
authors.affiliation: Shiseido Research Center, Hayabuchi, Kouhoku, Yokohama 224-8558, Japan. thirobe@nirs.go.jp
authors.forename: Tomohisa
authors.initials: T
authors.lastname: Hirobe
authors.forename: Rikako
authors.initials: R
authors.lastname: Furuya
authors.forename: Ohji
authors.initials: O
authors.lastname: Ifuku
authors.forename: Masatake
authors.initials: M
authors.lastname: Osawa
authors.forename: Shin-Ichi
authors.initials: S
authors.lastname: Nishikawa
mesh_terms: D000818:Animals; D000831:Animals, Newborn; D003994:Bucladesine / Q000494:pharmacology; D002454:Cell Differentiation / Q000187:drug effects*; D002455:Cell Division / Q000187:drug effects*; D002478:Cells, Cultured; D016895:Culture Media, Serum-Free; D004797:Enzyme-Linked Immunosorbent Assay; D000078404:Epidermal Cells; D016222:Fibroblast Growth Factor 2 / Q000494:pharmacology; D005434:Flow Cytometry; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000494:pharmacology*; D015603:Keratinocytes / Q000737:chemistry*; D008544:Melanocytes / Q000166:cytology / Q000187:drug effects*; D051379:Mice; D008810:Mice, Inbred C57BL
chemical_list: D016895:Culture Media, Serum-Free; D016222:Fibroblast Growth Factor 2; D003994:Bucladesine; D016178:Granulocyte-Macrophage Colony-Stimulating Factor