Effects Of Different Feeder Layers To Human Keratinocytes Proliferation In Vitro

At present, burn wounds are usually repaired by autologous free skin grafting in clinic practice. However, when to cure a patient with a large area skin burning, it is always short of autogenous healthy skin obtainable. With the development of tissue engineering, there was great improvement in human epidermic cells culture in vitro, and the rapid amplification of autogenous keratinocytes in vitro offered a new promising choice to cure burn wounds. But the period of keratinocyte amplification was about 2 weeks with usual culture method in which the 3T3 cells treated by 7 ray were taken as feeder layer and FAD was culture medium. This however, couldn't satisfy cure needs in time. In the present study, modification to traditional culture process was made to search for feeder layers more suitable to human keratinocytes proliferation and estabilish a more rapid procedure of human keratinocytes amplification in vitro.Feeder layers were prepared from 3T3 cells or primary cultured human fibroblasts, pre-treated by 6 Gry Co-60 α ray ormitomycin of different concentration ( 5 μ g / ml 10μg/ml 15 μ g / ml) . Epithelial cells were isolated from human fresh skin resected from operation patients and replaced on different prepared feeder layers. The growth state was observed.The findings were l)the feeder layer treated with mitomycin had the same function to accelerate the growth of keratinocytes as α ray exposure , 2)the concentration of 15μ g/ ml suitable toprimary fibroblasts and 10 g / ml suitable to 3T3 cells, and 3)the epithelial cells replaced on primary fibroblast feeder layer had a faster proliferation rate.So from the results of the study, we concluded that primary human fibroblast culture treated with mitomycin (15 g / ml) was an ideal feeder layer to human keratinocytes cultured in vitro.