人皮肤瘢痕成纤维细胞原代培养及其生长曲线的研究

目的 建立可靠的人皮肤瘢痕成纤维细胞培养方法,探索细胞的生长规律,为皮肤瘢痕体外研究提供平台.方法 采用组织块贴壁法原代培养瘢痕成纤维细胞,波形蛋白免疫细胞化学法鉴定成纤维细胞,CCK-8测定细胞生长曲线.结果 人皮肤瘢痕原代成纤维细胞培养成功,细胞形态为长梭形,波形蛋白表达阳性,16～18天可传第1代,以后每7～8天可传1代,传代后5～6天时为活跃期,第7天进入停滞期.结论 培养出的人皮肤瘢痕成纤维细胞在传代后第5～6天最适于于体外实验研究.

Study of primary culture of human skin scar fibroblasts and their growth curve

Objective To establish a reliable cultivation method for human skin scar fibroblasts and explore the growth pattern of the cells, and provide a plat for in vitro research of human skin scars. Methods Primary culture of human skin scar fibroblasts was performed with routine tissue block adhering to the plate wall, and the fibroblasts were identified by immunocytochemical staining of the vimentin, and cell growth curve was measured by CCK - 8. Results The primary culture of skin scar fibroblasts was successful. The fibroblasts have long - spindle shapes in cell morphology and positive vimentin expression. It takes 16 to 18 days for the first generation, and 7 to 8 days is required for each subculture. From the 5th to the 6th day after subculture, the cells were in the active period, while on the 7th day, the cells entered the period of stagnation. Conclusion Human skin scar fibroblasts can be cultured in stable condition in vitro, and the period from the 5th to 6th day after subculture is the most adequate time for in vitro experiments.