摘要
目的探索在体外培养条件下,转化生长因子(TGF)-β_1、胰岛素样生长因子(IGF)-1和骨形成蛋白(BMP)-2诱导间充质干细胞(MSCs)向软骨细胞分化的最佳组合。方法在体外单层培养条件下,利用TGF-β_1、IGF-I、BMP-2的组合作为诱导因子进行培养,用反转录聚合酶链反应(RT-PCR)检测细胞表达Ⅱ型前胶原mRNA水平,阿新蓝染色检测蛋白聚糖合成情况。结果BMP-2+IGF-I组、TGF-β组、TGF-β_1+IGF-I组、TGF-β_1+BMP-2组、TGF-β_1+IGF-I+BMP-2组Ⅱ型前胶原mRNA水平分别为0.147±0.062、0.478±0.069、0.489±0.022、0.653±0.013、0.807±0.051;而各组阿新蓝染色均阳性。结论TGF-β_1、IGF-I、BMP-2三者合用时诱导MSCs向软骨分化的效应最大。
Objective The present study aims to explore the optimal combination of growth factors, including TGF-β1, IGF-Ⅰ and BMP-2, for the induction of chondrogenesis of MSCs in vitro monolayer culture. Methods MSCs were induced to differentiate into chondrocytes in the medium with the following growth factors： TGF-β1, BMP-2 and IGF-Ⅰ, either alone or in combination. We detected two cartilage specific markers in differentiated cells. Alcian blue stain was used for aggrecan and RT-PCR for collagen type Ⅱ mRNA. Results The level of Collagen type II mRNA of group BMP-2＋IGF-Ⅰ,TGF-β1,TGF-β1＋IGF-Ⅰ,TGF-β1＋BMP-2 and TGF-β1＋IGF-Ⅰ＋BMP-2 was 0.147±0.062, 0.478±0.069, 0.489±0.022, 0.653±0.013, 0.807±0.051 respectively. Alcian blue stain in all groups were positive. Conclusion The TGF-β1＋IGF-Ⅰ＋BMP-2 group is the most effective combination in chondrogenesis induction.
出处
《中华风湿病学杂志》
CAS
CSCD
2007年第11期658-661,705,共5页
Chinese Journal of Rheumatology
