目的观察碱性成纤维细胞生长因子(bFGF)、胰岛素样生长因子(IGF)对体外培养骨髓基质干细胞(BMSc)增殖、分化的影响.方法 3～6月龄的雄性SD大鼠处死后,取双侧股骨、胫骨的骨髓进行体外培养.将含有不同细胞因子的培养基分为4组:①bFGF组;②IGF组;③bFGF+IGF组;④对照组.以上各组均含地塞米松、β-甘油磷酸钠、维生素C、青霉素、链霉素.显微镜下观察BMSc的生长特性,用MTT法测得的细胞相对值绘制生长曲线.采用改良Gomori氏钙钴法计数碱性磷酸酶(ALP)阳性率的细胞.结果 BMSc在传代培养前两天,各组的细胞数量无明显变化,从第二至第八天,bFGF组、IGF组、bFGF+IGF组的细胞数量均高于对照组,并且bFGF+IGF组明显高于其它3组.含细胞因子3组的ALP阳性率均高于对照组,bFGF+IGF组的阳性率最高.结论 bFGF、IGF都能促进BMSc增殖及向成骨细胞分化,并且两者同时运用优于单独使用;bFGF、IGF在促进BMSc增殖与分化时起协同作用.
Objective To study the effect of basic fibroblast growth factor (bFGF) and insulin-like growth factor (IGF) on proliferation and differentiation of bone mesenchymal stem cells (BMSc) in vitro culture. Methods The 3-6 months old male Sprague-Dawleg rats were killed, the marrow plugs obtained from bilateral femorae and tibiae were cultivated in vitro. The media were divided into four groups, according to contents of different cell factors.①bFGF group; ②IGF group; ③bFGF+ IGF group;④control group. Dexam-ethasone,β-glycerosphosphoric acid, vitamin C, penicillin, streptomycin were added in all above groups. The growth characteristics of BMSc were observed under the microscope, the relative value of the cells were measured and the growth curvature was drawn with MTT method. The ALPase positive cells were counted by the improved Gomori's method. Results At the beginning two days of culture, all of groups had no difference in cell quantity. From second to eighth day, the amount of cells in these three groups, including cell factors (bFGF, IGF,bFGF + IGF) were more than control group respectively, however, cells of bFGF+ IGF group were the most in all groups. Cells of the groups including cell factors expressed rate of ALPase positive cells were more than the cells of control group,moreover, the cells of bFGF+ IGF group which expressed the rate of ALPase cells were the most in all groups. Conclusion bFGF, IGF can enhance the proliferation and differentiation of BMSc into osteoblast,moreover, the effect of bFGF and IGF,added to media at the same time is better than the effect of bFGF and IGF added respectively. bFGF and IGF can promote the proliferation and differentiation of BMSc, with synergistic action.
China Journal of Orthopaedics and Traumatology