组织工程与重建外科杂志

组织工程与重建外科杂志 ›› 2014, Vol. 10 ›› Issue (3): 127-130.doi: 10.3969/j.issn.1673-0364.2014.03.002

• 论著 • 上一篇    下一篇

张力刺激皮肤成纤维细胞向肌成纤维细胞分化的实验研究

傅士博，温从吉，王琛，王丹茹   

  1. 上海交通大学医学院附属第九人民医院整复外科
  • 发布日期:2020-07-23

Myofibroblasts Differentiation of Skin Fibroblasts Induced by Mechanical Loading Based on 3D Culture in Vitro

FU Shibo,WEN Congji,WANG Chen,WANG Danru   

  1. Department of Plastic and Reconstructive Surgery,Shanghai Ninth People’s Hospital,Shanghai Jiaotong University School of Medicine;
  • Published:2020-07-23

摘要： 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。

关键词: 肌成纤维细胞, 机械张力, 三维培养

Abstract: 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.

Key words: Myofibroblasts, Mechanical loading, 3D culture

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