人皮肤角质形成细胞的分离与原代无血清培养

Isolation and serum-free culture of human skin primary keratinocytes

目的 :建立一种体外人皮肤角质形成细胞分离与原代培养的方法。方法 :采用两步消化法对皮肤进行消化 ,获取角质形成细胞进行体外无血清培养基培养 ,进行细胞形态学观察、免疫组织化学、透射电镜鉴定及生长曲线的绘制与分析。结果 :该方法可获取较多高纯度的角质形成细胞 ,且在体外可快速稳定增殖。结论

Objective To establish an optimal method for in vitro culture and amplification of primary keratinocytes from the human skin. Methods The keratinocytes were isolated from the human skin by two step combined dissociation.The cultured cells in serum free media were indentified by morphology, immunohistochemistry,and transmissive electronmicroscope. Growth curves were plotted and analyzed.Results A lot of highly pure keratinocytes could be obtained by this procedure and cultured in vitro steadily and rapidly. Conclusion Two step combined dissociation and serum free culture in vitro for human skin primary keratinocytes is an ideal method.