Isolation And Culture In Vitro Of Qinchuan-scalper Skin Cells

There are no establishment of qinchuan-scalper skin cell lines until 2002. The present experiment aimed to establish qinchuan-scalper skin cell lines. The culture method and characteristics of qinchuan-scalper skin fibroblast and epithelial cells were studied. The results as follows:1) The primary culture of qinchuan-scalper skin cells could be derived by fragment of tissue dispersed and cold digested single cell, Collegenase I (150IU mL-1') and trypsin(0.05%) being used at the same time is best to dissociate qinchuan-scalper skin tissue, Which is appropriate to obtain qinchuan-scalper skin cells.2) Hanks' buffer of trypsin not being added EDTA could obtain better the growth potential and the subsequent attachment of harvested skin cells t Then it being added EDTA have better digested potent.3) Dispased cold digested single cell is the best to obtain pured epithelial cell line; It is appropriate to obtain pured fibroblast cell line that repeated attachment and the pured method to their different sensibility to trypsin.4) Either low-sugar DMEM, TCM199, DMEM+F12(1:1) support qinchuan-scalper skin fibroblast cells growth well, but low-sugar DMEM caused best effect5) EOF and IGF-i added in low-sugar DMEM could promote qinchuan-scalper skin fibroblast cells growth potent respectively, yet they have cooperated effect each other.6) After passage, the qinchuan-scalper skin fibroblast cells were in lag phase in first two day, then entered log phase and persisted six days, they stayed at plateur phase for about three days; The mitotic index were most high at the fifth day after culture and toboggan at the sixth day. These showed qinchuan-scalper skin fibroblast cells were normal cells at four passages.7) Qinchuan-scalper skin fibroblast cells containted chromosome content at least the eighth passage.