bFGF基因转染对骨髓基质细胞生物学特性的影响

Changes of bioactivity of cultured bone marrow stromal cells transfected with basic fibroblast growth factor gene in vitro

目的:研究碱性成纤维细胞生长因子(bFGF)基因转染对骨髓基质细胞(BMSCs)生物学特性的影响,探讨细胞因子在骨缺损基因治疗的可行性,为进一步体内实验奠定基础。方法:利用脂质体将bFGF-pcDNA3重组真核表达载体导入体外培养的BMSCs中,对稳定表达bFGF的BMSCs从形态、增殖特性(MTT法)和细胞周期、碱性磷酸酶活性等生物学特性方面进行观察。采用SPSS10.0统计软件包进行t检验、方差分析,以P<0.05为差异有显著性。结果:转染bFGF-pcDNA3后,细胞生长曲线上移,倍增时间缩短,与未转染细胞相比,两者有显著差异(P<0.05),表明转染bFGF-pcDNA3后,细胞生长加快;细胞周期中增殖期细胞比例明显增大;而细胞形态和碱性磷酸酶活性无明显改变。结论:表达bFGF的BMSCs能促进自身的增殖,并不抑制细胞向成骨细胞方向分化。

PURPOSE： To study the changes of the bioactivity of cultured bone marrow stromal cells transfected with basic fibroblast growth factor gene in vitro,and to analyze the molecular mechanism of bFGF function. METHODS： bFGF-pcDNA3 gene was transfected into BMSCs by using Lipofect.Positive clones were selected with G418. The proliferativity （by MTT analysis） and the synthesis alkaline phosphatase （ALP）, the shape and cell cycle of the transfected BMSCs were examined. Student＇s t test and analysis of variances were used for statistical analysis with SPSS10.0. RESULTS： The growth curve of bFGF-pcDNA3 transfected cells was steeper than that of non-transfected cells, and statistic analysis confirmed that bFGF-pcDNA3 transfected ceils growed more quickly（P〈O.05）. There was significant difference in cell cycle, whereas the shape and the alkaline phosphatase （ALP） activity remained unchanged. CONCLUSION： The proliferation content of genetically modified BMSCs were increased significantly.