Abstract:

Investigation of the culture methods of human keratinocytes in vitro Bu Xiao-lin, Lu hongguang （Department of Dermatology, Affiliated hospital,Guiyang Medical College, Guiyang 550004, China） Abstract: Objective To compare the biological characteristics of human keratinocytes (KC) in keratinocyte serum-free medium(K-SFM) and defined keratinocyte serum-free medium(DK-SFM).Methods The keratinocytes were harvested from foreskin. A single cell suspension were dissociated by Dispase and trypsin and cultured respectively in the medium of K-SFM and DK-SFM. The morphology of KC was observed under light microscope. The cell cycle and expression of epidermal growth factor receptor (EGFR) were determined with flow cytometry.Result KC grew fast and the numbers of cells and colony appeared much more signifcant high in K-SFM than in DK-SFM. The cells in K-SFM could be maintained in culture up to 4 passages but couldn’t pass into the second generation in DK-SFM. There were also higher expression of EGFR were recorded in the cells of K-SFM than in DK-SFM. Conclusion In contrast to DK-SFM, K-SFM seem to take advantage of the cultivation of human KC in vitro.

Key words: keratinocytes;serum-free medium;culture method