【摘要】 目的:研究碱性成纤维细胞生长因子(bFGF)和骨形成蛋白-2(BMP-2)联合应用对体外培养兔骨髓间充质干细胞(BMSCs)增殖与骨向分化的影响。方法:体外培养兔骨髓间充质干细胞,在第2代细胞培养液中加入不同浓度的bFGF和BMP-2,依据加入bFGF和BMP-2浓度的不同分为5个实验组(组1:80 ng/ml bFGF;组2:80 ng/ml BMP-2;组3:30 ng/ml bFGF+30 ng/ml BMP-2;组4:50 ng/ml bFGF+50ng/ml BMP-2;组5:80 ng/ml bFGF+80 ng/ml BMP-2)和对照组(不加任何生长因子),采用绘制生长曲线,四唑盐比色法(MTT),碱性磷酸酶(ALP)活性检测法和碱性磷酸酶(ALP)免疫组化染色法比较各组间差异,观察不同浓度的bFGF和BMP-2联合应用对免骨髓间充质干细胞增殖与骨向分化的影响。结果:与对照组相比,单独应用80 ng/ml bFGF可显著促进BMSCs的增殖,但对BMSCs的骨向分化显著抑制;单独应用80 ng/ml BMP-2对BMSCs的增殖和骨向分化均有促进作用;30 ng/ml bFGF+30 ng/ml BMP-2、50 ng/ml bFGF+50 ng/ml BMP-2和80 ng/ml bFGF+80 ng/ml BMP-2可显著地促进BMSCs增殖和促进BMSCs的骨向分化,且呈正性剂量-效应关系,联合应用两种生长因子较二者单独应用促细胞增殖及骨向分化的效果更为显著。结论:一定浓度范围内,bFGF和BMP-2的联合应用促进BMSCs增殖的同时也促进其骨向分化,两者对BMSCs有明显的协同增强的生物学效应。更多还原

【Abstract】 Objective To study the effects of combination of basic fibroblast growth factor(bFGF)and bone morphogenetic protein-2(BMP-2)on proliferation and osteogenic differentiation of rabbit bone mesenchymal stem cells(BMSCs)in vitro culture. Methods BMSCs were cultured in vitro,different concentrations of bFGF and BMP-2 were added into culture fluid of the 2nd passage. According to the concentration ofbFGF and BMP-2,five experimental groups were entered(group1:80 ng/ml bFGF;group2:80 ng/ml BMP-2;group3:30 ng/ml bFGF+30 ng/ml BMP-2;group4:50 ng/ml bFGF+50 ng/ml BMP-2;group5:80 ng/ml bFGF+80 ng/ml BMP-2), no growth factor was added into the culture fluid in control.The growth curve drawing,tetrazolium salts chromatometry(MTT),ALP dynamics method and immunohistochemitry staining were used to compare the difference between the groups and to observe the effects of combination of bFGF and BMP-2 on proliferation and osteogenic differentiation of BMSCs.Results Compared with the control group, 80 ng/ml bFGF could contribute to the proliferation of BMSCs significantly,but had inhibition effects on the osteogenic differentiation of BMSCs;80 ng/ml BMP-2 had promotive effects both on the proliferation and osteogenic differentiation of BMSCs;30 ng/ml bFGF+30ng/ml BMP-2,50 ng/ml bFGF+50ng/ml BMP-2 and 80 ng/ml bFGF+80 ng/ml BMP-2 not only promoted BMSCs proliferation significantly,but also promoted the osteogenic differentiation of BMSCs,showing positive dose-effect.Conclusion In certain concentration range,BMP-2 in combination with bFGF can promote the proliferation and osteogenic differentiation of BMSCs.更多还原