优化贴壁法人皮肤成纤维细胞的原代培养

目的:优化贴壁法原代培养人皮肤成纤维细胞的条件,建立稳定?经济?高效可行的人皮肤成纤维细胞培养方法?方法:采用包皮环切术后皮肤组织,剪成均匀组织块后贴壁,皮肤边缘长出薄层致密细胞团后胰酶消化?传代,并对所培养的细胞进行形态学观察及细胞免疫荧光鉴定?此方法与酶消化法相对照?结果:优化贴壁法成功率100%,取得组织后2 d沿皮肤边缘均开始生长高密度类圆形细胞层,在1周内可获得高质量?高数量的细胞,传代贴壁后经镜下形态观察及细胞免疫荧光染色确定为皮肤成纤维细胞?结论:较之酶消化法,优化贴壁法细胞存活率高,活性及纯度好?该方法可稳定?经济地获得足够数量的细胞?

Optimized Primary Culture of Human Skin Fibroblasts by Tissue Adherent Method

ABSTRACT] Objective:To optimize and establish a stable, economical and feasible protocol for human skin fibroblasts primary culture. Methods:Foreskins were minced into pieces and placed into gelatin-coated 6-well plates. When there were lumpy tissues coming out, cells were digested and passaged. They were then observed by morphology and immunochemistry. This method was contrasted with enzyme digestion procedure. Results:The success rate of optimized tissue adherent method was 100%. Lumpy tissues consisted of High density cells grew from the edge of skin tissues within 2 days. High numbers of viable cells which were identified as fibroblasts by morphology and immunochemistry can be obtained within a week. Conclusion:Cells cultured by optimized tissue adherent method are with high survival rate, activity and purity. This method is more stable, economical and feasible in contrast with enzyme digestion procedure.