A cloned, immortal line of murine melanoblasts inducible to differentiate to melanocytes.
Cultures of differentiated melanocytes can readily be grown from the dissociated epidermis of neonatal mice, and immortal cell lines often develop from these. However, the first cells that grow and transiently dominate the cultures, while similar to melanocytes, are unpigmented. These have been shown to be precursors of melanocytes and may be termed melanoblasts. Under our previous standard culture conditions, involving the use of keratinocyte feeder cells, foetal calf serum, the phorbol ester 12-O-tetradecanoyl phorbol acetate (TPA) and cholera toxin, all the melanoblasts spontaneously differentiated to pigmented melanocytes within about 3 weeks. We now describe some factors affecting the proliferation and differentiation of diploid murine melanoblasts in the presence of serum. Murine stem cell factor/steel factor (SCF), basic fibroblast growth factor (bFGF) and murine leukaemia inhibitory factor/differentiation-inhibiting activity (LIF/DIA) all increased melanoblast numbers. SCF and LIF also slightly inhibited melanoblast differentiation, while cholera toxin and TPA promoted differentiation. Using some of these findings, and by regular replacement of keratinocyte or fibroblastoid feeder cells, we have established a clonal line of immortal murine melanoblasts, 'melb-a'. These cells express tyrosinase-related protein-2 but not, in general, tyrosinase. They can be induced to differentiate irreversibly to functional melanocytes (also proliferative and immortal) by plating in the absence of feeder cells. Thus a new immortal melanocyte line, 'melan-a2', has also been produced.
Sviderskaya
E V
EV
St George's Hospital Medical School, London, UK.
Wakeling
W F
WF
Bennett
D C
DC
D000818:Animals; D015815:Cell Adhesion Molecules / Q000494:pharmacology; D002452:Cell Count; D002454:Cell Differentiation / Q000187:drug effects; D002455:Cell Division; D002460:Cell Line; D002772:Cholera Toxin / Q000494:pharmacology; D002999:Clone Cells; D000078404:Epidermal Cells; D004817:Epidermis / Q000201:enzymology; D016222:Fibroblast Growth Factor 2 / Q000494:pharmacology; D006131:Growth Inhibitors / Q000494:pharmacology; D016298:Hematopoietic Cell Growth Factors / Q000494:pharmacology; D006651:Histocytochemistry; D007150:Immunohistochemistry; D015850:Interleukin-6; D019746:Intramolecular Oxidoreductases; D007535:Isomerases / Q000378:metabolism; D053605:Leukemia Inhibitory Factor; D008222:Lymphokines / Q000494:pharmacology; D008544:Melanocytes / Q000166:cytology* / Q000201:enzymology; D051379:Mice; D008810:Mice, Inbred C57BL; D019089:Stem Cell Factor; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology
D015815:Cell Adhesion Molecules; D006131:Growth Inhibitors; D016298:Hematopoietic Cell Growth Factors; D015850:Interleukin-6; D053605:Leukemia Inhibitory Factor; C507838:Lif protein, mouse; D008222:Lymphokines; D019089:Stem Cell Factor; D016222:Fibroblast Growth Factor 2; D002772:Cholera Toxin; D007535:Isomerases; D019746:Intramolecular Oxidoreductases; C042501:dopachrome isomerase; D013755:Tetradecanoylphorbol Acetate