胰岛素样生长因子-1基因转染脂肪间充质干细胞向软骨细胞分化的研究

Experimental study on chondrocyte-differentiation of adipose mesenchymal stem cells by gene transfection of insulinlike growth factor-1 in vitro

目的观察胰岛素样生长因子（IGF）-1基因转染的脂肪间充质干细胞（ADSCs）向软骨细胞分化的效果。方法原代培养兔ADSCs，免疫荧光法检测细胞表面抗原CD44、CD49；脂质体介导人IGF-1基因转染兔ADSCs联合低浓度血清培养基向软骨细胞分化诱导，RT-PCR及Western blot方法检测IGF-1的表达，MMT法绘制细胞增殖曲线、甲苯胺蓝染色软骨结节、免疫组织化学检测Ⅱ型胶原的表达。结果脂肪间充质干细胞CD44、CD109表达阳性，基因转染后细胞IGF-1表达阳性，细胞增殖速度增快，出现软骨结节，Ⅱ型胶原表达增高。结论从脂肪组织中能够分离出增殖旺盛的ADSCs，IGF-1在ADSCs内获得稳定表达，细胞增殖能力增强，促进其向软骨细胞分化。

Objective To explore the biologic activity of adipose mesenchymal stem cells （ ADSCs） and the effect of IGF-1 gene in transforming ADSCs into chondrocytes in vitro. Methods Primary culture of the ADSCs from fat tissue was done. CD44 and CD109 were detected by immunofluorescence. ADSCs were transfected with the method of stable transfection of gene IGF-1 with Liprofectamine 2000 and the low concentration blood serum culture-medium. The expression of IGF-1 was detected by the method of RT-PCR and Western blot. The proliferation of ADSCs was assayed MTT. The cartilage nodus was analyzed by toluidine blue stain. The expression of collagen Ⅱ was tested by immtmofluorescence. Results ADSCs were positive for CD44 and CD109. After gene transfection, the ADSCs expressed IGF-1, the proliferation of ADSCs was increased,cartilage nodus was seen and the expression of collagen Ⅱ up-regulated. Conclusion Exogenous IGF-1 was stably expressed in ADSCs. Tranfection of IGF-1 into ADSCs increased their proliferation and promote the differentiation into chondrocytes in vitro.