含AB型血清培养体系人角质形成细胞培养

目的：探讨人体角质形成细胞的培养技术，观察人角质形成细胞生长的过程，为培养角质形成细胞膜片提供新的方法。方法：采用改良后的含胎牛和人AB型血清培养体系进行人角质形成细胞的培养。结果：含人AB型血清培养体系培养的角质形成细胞5d融合达70％，9d达90％，11d完全融合，细胞分裂指数高峰在9d左右，可达1．1％，而不合AB型血清组培养的细胞5d后即出现退化样生长，不形成细胞膜片。进一步对含A13型血清的不同培养系统进行培养，将增殖高峰期培养的角质形成细胞应用MTT比色法测细胞增殖活性，进行比较，P〉0．05，表明添加F12、TC199、1640后对培养的角质形成细胞增殖作用无差异。结论：人AB型血清能够对体外培养的角质形成细胞生长起到刺激作用。

THE CULTIVATION OF HUMAN KERATINOCYTES IN CULTURE SYSTEM WITH AB TYPE OF HUMAN SERUM

Objective:To explore the cultivation technique of human keratinocytes in vitro and provide a new method for cultivation of keratinocytes lamina by observing the growth of human keratinocytes in the different culture system.Methods:Modified culture system with fetus calf and/or AB type of human serum was used for culturing human keratinocytes.Results:The cultivated keratinocytes in culture system with AB type of human serum expanded to 70% of total area in 5 days,to 90% in 9 days,and to 100% in 11days.The mitotic index reached to the peak value(1.1%) in about 9 days.Five days later,the cultivated cells were degenerated in culture system without AB type of human serum,and the lamina couldn't develop.Furthermore,the cells were cultured in different culture system with AB type of human serum.Proliferation activity of the keratinocytes at the stage of proliferating peak was measured with MTT colorimetry.The results showed no difference(P>0.05) in the proliferation of cultured keratinocytes in different culture system with F12,TC199 and 1640 media respectively.Conclusion:AB type of human serum could stimulate the growth of human keratinocytes in vitro culture system.