bFGF真核表达质粒的构建及骨髓基质细胞转染

Construction of eukaryotic expression vectors of basic fibroblast growth factor and transfection in rabbit bone marrow stromal cells

目的探讨bFGF基因转染骨髓基质干细胞的方法及可行性。方法构建bFGF基因真核表达质粒,用脂质体法介导转染骨髓基质细胞,通过形态学观察、免疫组化、ELISA法、及RT-PCR方法检测bFGF基因转染骨髓基质干细胞的成功性,以及转染后骨髓基质干细胞的生物学特性。结果bFGF基因成功转染骨髓基质干细胞,并能持续稳定的分泌bFGF蛋白,可诱导骨髓基质干细胞向成软骨方向分化。结论bFGF基因可以转染骨髓基质干细胞,并能诱导骨髓基质干细胞向成软骨方向分化。

Objective To investigate approach and possibility of transferring basic fibroblast growth factor （bFGF） gene into rabbit bone marrow stromal cells （BMSCs）. Methods The eukaryotic expression vectors harboring bFGF cDNA were constructed and transfected into rabbit BMSCs mediated by liposome. The transcription and expression ofbFGF gene in the transfected BMSCs were detected by means of morphological observation, immunohistochemistry, enzyme-linked immunosorbent assay （ELISA） and RT-PCR. The changes in the biological characteristicS of the transfected MSCs were also observed. Results Stable overexpression of bFGF protein was detected in the transfected BMSCs, which showed differentiation towards chondrocyte linage. Conclusion Stable expression ofbFGF gene in transfected BMSCs can induce cell differentiation into chondrocyte lineage.