生长分化因子5对大鼠骨髓间质干细胞生长分化的影响

The effect of GDF5 on the proliferation and differentiation of bone marrow stromal cells

目的研究生长分化因子5(growth and differentiation 5,GDF5)对大鼠骨髓间质干细胞(bone marrow mesenchymal stem cells,BMSCs)生长分化的影响。方法分离和培养大鼠BMSCs,培养液添加GDF5(添加浓度分别为0、10、100ng/mL),用细胞计数、MTT比色检测细胞增殖变化,用甲苯胺蓝、阿辛蓝染色和RT-PCR检测细胞蛋白多糖(PG)和Ⅱ型胶原合成。结果在低糖DMEM中,GDF5组细胞数量显著增多,可以促进BMSCs生长。在内含地塞米松、胰岛素等的DMEM/F12成软骨诱导液中,阿辛蓝浓度OD值在GDF5组均随时间增加而增高,Ⅱ型胶原合成增多可以诱导BMSCs向软骨细胞分化。结论 GDF5在低糖DMEM中可促进BMSCs的增殖;在成软骨诱导体系中,可使单层培养的BMSCs聚集诱导其向软骨细胞分化,并形成软骨小结。

Objective To explore the effect of GDF5 on the differentiation of bone marrow stromal cells （BMSCs） into chondrocytes. Methods The methods of harvest and culture of BMSCs from rats were used; GDF5 was added in culture fluid, cell counting and MTT were used to test the cell proliferation; Toludine blue, alcian blue staining and RT-PCR were used to examine the synthesis of proteoglycan and collagen. Results GDF5 was showed to be promotor for BMSCS growth in low sugar DMEM＇s fluid, resulting in that the number of BMSCs increased, logarithmic growth was in advance, cell aggregated followed the confluence, the absorbed optical density on cell growth enhanced. In the DMEM/F12 cultured fluid to induce the chondrogensis contained Dexamethasone, Insuline and so on, the effect of GDF5 was showed to induce the differentiation of BMSCs into chondroeytes. Conclusion GDF5 could promote the proliferation of BMSCs in the low sugar DMEM＇s fluid, make the aggregation of BMSCs in the system induced to chondrogensis and induce the BMSCs differentiation into chondrocyte and form the cartilage nodule.