CAJ | 학술논문

目的：探索机械张力刺激诱导基于三维培养的皮肤成纤维细胞向肌成纤维细胞分化的可行性。方法将正常人皮肤成纤维细胞三维培养于Ⅰ型鼠尾胶原支架中，通过机械张力装置进行拉伸实验（牵拉组，n=9），并设立对照（非牵拉组，n=9），绘制细胞生长曲线，免疫荧光染色和实时定量PCR检测成纤维细胞的分化程度。结果三维培养环境下细胞增殖缓慢，牵拉组有促进细胞增殖的趋势，但与非牵拉组相比，统计学差异不显著；免疫荧光染色显示，牵拉组成纤维细胞α-SMA和fibronectin染色亮度明显高于非牵拉组；实时定量PCR检测显示，牵拉组α-SMA、纤连蛋白和Ⅰ、Ⅲ型胶原mRNA表达高于非牵拉组。结论机械张力可成功诱导基于三维培养的皮肤成纤维细胞向肌成纤维细胞分化。
목적：탐색궤계장력자격유도기우삼유배양적피부성섬유세포향기성섬유세포분화적가행성。방법장정상인피부성섬유세포삼유배양우Ⅰ형서미효원지가중，통과궤계장력장치진행랍신실험（견랍조，n=9），병설립대조（비견랍조，n=9），회제세포생장곡선，면역형광염색화실시정량PCR검측성섬유세포적분화정도。결과삼유배양배경하세포증식완만，견랍조유촉진세포증식적추세，단여비견랍조상비，통계학차이불현저；면역형광염색현시，견랍조성섬유세포α-SMA화fibronectin염색량도명현고우비견랍조；실시정량PCR검측현시，견랍조α-SMA、섬련단백화Ⅰ、Ⅲ형효원mRNA표체고우비견랍조。결론궤계장력가성공유도기우삼유배양적피부성섬유세포향기성섬유세포분화。
Objective To explore the feasibility of inducing myofibroblasts differentiation of skin fibroblasts by mechanical loading based on three dimensional culture in vitro. Methods Human dermal fibroblasts were isolated and three dimensionally cultured in type I rat tail collagen scaffold. Then mechanical loading were added by stretching the gel (stretch group, n=9), and control group was set up (non-stretch group, n=9). The situation of myofibroblasts differentiation were tested by drawing cell growth curve, immunofluorescent staining and Real-Time PCR. Results Cell proliferation was slowed down under mechanical loading environment. The cell proliferation was promoted in stretch group, but there was no significant difference comparing with non-stretch group. Dyeing brightness of α-SMA and fibronectin were significantly higher in stretch group showed by immunofluorescent staining. Higher mRNA expression of α-SMA, fibronectin, type I and III collagen genes were observed in stretch group by real time PCR. Conclusion The model of mechanical loading inducing myofibroblasts differentiation of skin fibroblasts based on 3D culture could be successfully established.