关键词: 黑素细胞, 细胞培养, 纯化, 包被

Abstract: Objective:To compare the influence of different coating materials and cultural conditionson the purification and growth of human epidermal melanocytes.Methods:The full-thinckforeskin, epidermis and cell suspension obtained from human foreskin were cultured in the plates which were precoated with Matrigel or laminin respectively. When having reached 80–90% confluence, the cells were treated with 0.05% trypsin-EDTA for 4 minutes and resuspendedin M254 medium which were supplemented with G418 and 5-BrdU, respectively. Whereafter, the purity of melanocytes was observed by animmunofluorescence staining with melanocytes’ markers. Results: During the primary culture, the cells’suspension generated more cells at faster speed comparing with skin explants and epidermal specimen. Moreover, the epidermis released cells earlier and proliferated quickly over skin explants. The melanocytes in the plates coated with laminin other than with Matrigel displayed faster and better growth. The unwanted keratinocytes and fibroblasts were removed by using differentedtrypsinition combining with supplement of G418 or 5-BrdU.Conclusions: Using a plate coated with laminin to culture cell suspension from human foreskin, and via a differentedtrypsinization combining with supplement of small doses of G418 to subculture the cells, is admelavantageous to the melanocyte purification, without affecting their growth.

Key words: melanocytes, culture, purification, coat.