Purification of epidermal melanocytesin culture
10.11958/20161212
- VernacularTitle:表皮黑素细胞的纯化
- Author:
Di LI
;
Ruzhi ZHANG
- Keywords:
melanocytes;
cell culture techniques;
purification;
coating materials
- From:
Tianjin Medical Journal
2017;45(1):5-8
- CountryChina
- Language:Chinese
-
Abstract:
Objective To compare the influence of different coating materials and cultural conditions on the purification and growth of human epidermal melanocytes. Methods The full-thick foreskin, 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 resuspended in M254 medium, which were supplemented with G418 and 5-BrdU, respectively. The purity of melanocytes was observed by an immunofluorescence staining with melanocyte markers. Results During the primary culture, the cell suspension generated more cells at faster speed compared with those of 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 differentiation trypsinition combined with supplement of G418 or 5-BrdU. Conclusion Using a plate coated with laminin to culture cell suspension from human foreskin, and via a differentiation trypsinization combined with supplement of small doses of G418 to subculture the cells, is advantageous to the melanocyte purification, without affecting their growth.
