【摘要】 目的:建立人皮肤成纤维细胞的体外原代及传代培养并鉴定的技术方法。方法:取临床无菌切除的幼儿包皮,利用胶原酶分离表皮和真皮,将真皮剪成肉糜状,再用胰蛋白酶消化,接种于培养皿,加少许含10%小牛血清的DMEM-高糖培养液进行培养,次日补加培养液。原代长满后,进行传代,并对所培养的细胞进行形态学观察及组化鉴定。结果:接种次日倒置镜下可见有长梭形细胞迁出、生长,5-6d进行传代,传代后约5d长满,可长期传代。免疫组化Vimentin表达阳性。结论:该方法可快速高效获得构建组织皮肤真皮所需的成纤维细胞。更多还原

【Abstract】 Objective:To explore the method of isolation,cultivation and identification of human skin fibroblasts in vitro.Methods:By digesting child foreskin with collagenase typeⅡ to isolate epidermis and corium.The shearing corium was digesting with trypsin.Dulbecco′s modified Eagle′s medium(DMEM)was the basic culture medium,being supplemented with 10%bovine calf serum.Cellular morphologies were observed by inverted phase contrast microscopy.The cultured cells were identified by vimentin immunohistochemical.Results:The spindle shaped fibroblasts could find by inverted phase contrast microscopy in second day.The fibroblasts could passage in fifth day.And immunohistochemical of vimentin was positive in cultured fibroblasts.Conclusion:It is an effective method to cultivate fibroblasts with high productivity and survival rate.更多还原