表皮细胞培养条件的优化  

Effects of EGF in serum-free medium on primaryculture of epidermal cells.

作　　者：孙曙光[1] 程大胜[2] 王良喜[1] 王静[1] 毛学飞[1] 曹淑琴[1] 

机构地区：[1]解放军第97医院,江苏徐州221004 [2]第二军医大学第一附属医院

出　　处：《山东医药》2006年第5期13-14,共2页Shandong Medical Journal

基　　金：南京军区医药卫生科研基金资助课题(2002-09-26)

摘　　要：目的探讨生长因子（EGF）在人表皮细胞无血清培养中的作用，寻找较好组合．以期加快表皮细胞成皮。方法选择EGF、胰岛素、霍乱毒素、氢化考的松4个实验因素，每个因素设立4个浓度水平。利用L16（4X4）正交表设计试验，考察不同培养条件对表皮细胞生长情况、克隆形成率的影响。结果EGF在无血清培养中对培养人表皮细胞的生长及增殖有明显促进作用（P〈0．01），在一定范围内（5～20ng／m1）随EGF浓度提高，效应加强；胰岛素浓度在5g／ml时对培养细胞的生长及增殖影响最大；霍乱毒素浓度在4．2ng／ml时作用最好；氢化考的松作用不明显。EGF、胰岛素、霍乱毒素、氢化考的松分别取20ng／ml、5μg／ml、8．4ng／ml、0．2μg／ml浓度时细胞生长及增殖效果最好。结论采用此优化的培养方法，表皮细胞的生长比较迅速，分化不太明显，可保持较旺盛的增殖力。Objective： To investigate the effects of several factors in serum-free medium on primarY culture of epidermal cells, try to find an optimal formula to promote growth and proliferation of epidermal cells. Methods Epidermal cells were isolated and primary cultured in serum-free medium with different serum substitute. The orthogonal trial of L16（44） was used to investigate the effects of four factors （epidermal growth faetor, insulin, cholera toxin, and hydrocortisone at four different concentrations） on the growth of viable cells and colony forming efficiency （CFE） of the isolated epidermal cells. Results： ①Epidermal growth factor （EGF） had significant effect on promoting growth and proliferation of epidermal cells. In the certain concentration range （5- 20ng/ml）, the higher the concentration, the stronger the effects. ②At a concentration of 5μg/ml, insulin had the maximum effects on the growth and proliferation of epidermal cells, For cholera toxin, a concentration of 4. 2 ng/ml exerted the effect. ③In the concentration range investigated, hydrocortisone did not show any significant effects on the growth and proliferation of epidermal cells, In primary culture, an optimal formula of the four factors contains 20ng/ml EGF, 5g/ml insulin, 8.4ng/ml cholera toxin, and 0.2g/ml hydrocortisone. Conclusions： Epidermal cells growth rapidly with a slight differentiafion,and can keep an active proliferation by using the optimal formula.

关 键 词：表皮细胞培养 克隆 正交表 

分 类 号：R644[医药卫生—外科学]