Selective growth and serial passage of mouse melanocytes from neonatal epidermis in a medium supplemented with bovine pituitary extract.
Suspensions of disaggregated epidermal cells from skins of newborn C57BL/10JHir mice were plated in a growth medium that consisted of Ham's F-10 plus bovine pituitary extract (BPE), insulin, and transferrin. Fetal bovine serum (FBS) was added to the culture medium at a concentration of 4% at the time of plating. On the second day of culture, a small number of melanocytes was randomly distributed among large sheets of keratinocytes. From the third day onward, FBS was excluded from the culture medium to prevent the proliferation of keratinocytes and fibroblasts. The melanocytes began to grow preferentially, and after 12 days pure and enriched populations of melanocytes could be harvested. In the absence of the proliferation of keratinocytes and fibroblasts, melanocytes could be serially passaged in the growth medium supplemented with a conditioned medium (CM) prepared from keratinocyte-enriched cultures, namely, those at the early stages of the primary culture. FBS was added at a concentration of 1% for the first day. These results suggest that both BPE and keratinocyte CM contain growth factors required for proliferation of melanocytes.
Hirobe
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Division of Biology, National Institute of Radiological Sciences, Chiba, Japan.
D000818:Animals; D002455:Cell Division; D002470:Cell Survival; D002478:Cells, Cultured; D003470:Culture Media; D000078404:Epidermal Cells; D006133:Growth Substances / Q000494:pharmacology; D007700:Kinetics; D007980:Levodopa / Q000494:pharmacology; D008544:Melanocytes / Q000166:cytology* / Q000187:drug effects; D051379:Mice; D008810:Mice, Inbred C57BL; D010902:Pituitary Gland; D012692:Serial Passage; D014020:Tissue Extracts / Q000494:pharmacology
D003470:Culture Media; D006133:Growth Substances; D014020:Tissue Extracts; D007980:Levodopa