负载转化生长因子β1的大鼠脂肪干细胞成软骨细胞分化

Differentiation of rat’s ADSCs loaded by TGF-β1 Gene to chondrocytes

目的转化生长因子β1基因转染大鼠的脂肪基质干细胞(ADSCs),诱导其向软骨细胞分化,为软骨组织工程学种子细胞提供新方法。方法分离、培养大鼠的ADSCs;采用免疫荧光法进行鉴定;TGF-β1基因转染ADSCs,对转染后细胞进行筛选;MTT法测定筛选后细胞的增殖活性;RT-PCR、Western blot对筛选后细胞的表达进行检测。结果成功分离、培养大鼠的ADSCs;细胞表面标志CD29和CD44表达阳性,CD106和CD34表达阴性;基因转染后细胞的增殖力变强;转染转化生长因子β1的ADSCs在目的基因的表达和软骨特异性基质的分泌上明显高于转染空载体组和对照组;结论转化生长因子β1基因转染后ADSCs具有了软骨细胞的表型特征,可以用作组织工程学的种子细胞。

[Objective] To induce the differentiation of rat＇s adipose stromal cells（ADSCs） into chondrocytes by transfecting ADSCs with TGF-β1 gene so as to provide a new method for tissue-engineering cartilage seed cells. [Methods] Rat＇s ADSCs were isolated and cultured, then the cells were identified by immune fluorescent assay. After being transfected the with TGF-β1, the cells were selected with transgene. MTT was used to detect the proliferation of the selected cells. Meanwhile, RT-PCR, and Western blot were used to analyze the expression level of the selected cells. [Results] Rat＇s ADSCs were successfully separated and cultured. Cell surface markers CD29 and CD44 were positive, whereas CD106 and CD34 were negative. The proliferative ability became stronger after being transfected with TGF-β1 gene. The transfected group had higher destined gene protein expression and cartilage specific matrix secretion than the vector group and control group. [Conclusion] Transfected ADSCs with TGF-β1 gene have phenol-features of chodrocytes, and can be used as the tissue-engineering seed cells.