bFGF重组慢病毒对羊骨髓间充质干细胞成骨功能影响的研究

Effect of bFGF Recombinant Lentiviral Vector on the Osteogenic Potential of Goat Bone Marrow-derived Mesenchymal Stem Cells

目的:探讨碱性成纤维生长因子(basic Fibroblast Growth Factor,bFGF)基因转染对体外培养的骨髓间充质干细胞(Bone Marrow mesenchymal Stem Cells,BMSCs)的增殖和骨向分化的影响,为构建组织工程骨的种子细胞做理论依据。方法:构建携带人bFGF基因的慢病毒载体,转染诱导后羊的BMSCs,得到bFGF转染组,选择未转染的BMSCs为对照组。两组细胞采用实时定量PCR技术和蛋白质印迹法技术检测Collagen-Ⅰ、OC、OPN等成骨相关基因的mRNA水平及蛋白水平相对表达量的变化情况。结果:bFGF组OPN和Collagen-Ⅰ基因mRNA水平表达量较对照组增高,差异有统计学意义(P<0.05),两组细胞在OC基因mRNA水平表达量上差异无统计学意义;bFGF组细胞在OPN、OC和Collagen-Ⅰ基因蛋白水平上表达量均高于对照组,且差异有统计学意义。结论:bFGF基因转染能增强羊BMSCs增殖和骨向分化的能力,可作为组织工程骨的种子细胞。

Objective：To study the effects of bFGF transfection on the proliferation and osteogenic differentiation of bone marrow mesenchymal stem cells.Methods：Lentiviral vectors carrying bFGF were constructed to transfect sheep bone marrow mesenchymal stem cells.Cells were divided into two groups：bFGF group,and control group（non-transfected goat bone marrow mesenchymal stem cells）.Real-time quantitative PCR and Western-blot were used to detect the changes of mRNA levels and protein levels of collagen-I,osteocalcin and osteopontin.Results：Expressions of collagen-I and osteopontin in mRNA levels were higher in transfection group than those in the control group,and the difference was statistically significant（P〈0.05）,But the expressions of osteocalcin in mRNA levels were not significant different in two group.Expression of OPN,Collagen- I,and OC of transfection group was higher than that in the control group,and the difference was statistically significant.Conclusion：bFGF transfection can enhance the ability of bone marrow mesenchymal stem cells on proliferation and osteogenic differentiation,and they are suitable for seed cells for bone tissue engineering.