原代培养人增生性瘢痕成纤维细胞的生物学行为

背景：增生性瘢痕组织的形成是创面愈合过程中不可避免的,但是增生性瘢痕组织的形成产生很多不良的影响。目的：建立可靠的人增生性瘢痕成纤维细胞的原代培养方法。方法：采用组织贴壁法和消化法分别进行人皮肤增生性瘢痕成纤维细胞的原代培养,使用含体积分数10%胎牛血清的DMEM培养基,37 ℃,体积分数5%CO2,饱和湿度下培养,分别描述其生长曲线、形态及波形蛋白的表达。结果与结论：组织贴壁法人皮肤增生性瘢痕成纤维细胞培养成功,20-40 d可传第1代,以后每7-10 d可传1代,细胞为长梭形,波形蛋白表达阳性;消化法培养人皮肤增生性瘢痕成纤维细胞成功,15-20 d细胞可融合成片,细胞为长梭形,波形蛋白表达阳性。进一步证实两种方法进行人增生性瘢痕成纤维细胞的原代培养均培养成功。

Primary culture of human hypertrophic scar fibroblasts and its biological behavior

BACKGROUND:Hypertrophic scar is inevitable in the process of wound healing, but the hypertrophic scar tissue can result in many adverse effects.OBJECTIVE:To establish a reliable method to culture human hypertrophic scar fibroblasts in vitro.METHODS:Human hypertrophic scar fibroblasts were cultured primarily in Dulbecco’s modified Eagle’s medium containing 10% fetal bovine serum using tissue adherent method and digestion method. Fibroblasts were incubated at 37℃in a humidified atmosphere of 5% carbon dioxide. Cellular morphologies were observed, the growth curve was drawn, and vimentin expression wass detected.RESULTS AND CONCLUSION:(1) After cultured with tissue adherent method, human hypertrophic scar fibroblasts were cultured successfully, first passaged within 20-40 days and then continued to passage every 7-10 days. The cells were long spindle-shaped, and vimentin was positive. (2) Human hypertrophic scar fibroblasts, cultured using digestion method, were cultured successfully and confluent at 15-20 days. The cells were long spindle-shaped, and vimentin was positive. These data have confirmed that both tissue adherent method and digestion method are successful in the primary culture of human hypertrophic scar fibroblasts.