不同基因转染对骨髓基质干细胞成骨活性的影响

为了优化骨组织工程种子细胞，探讨不同基因对骨髓基质干细胞（MSCs）成骨活性的影响，我们首先利用RT—PCR方法获得了全长BMP-2．VEGF 165及bFGF基因，克隆入真核表达载体peDNA3．0，鉴定正确后，经脂质体介导转入分离培养的大鼠骨髓基质干细胞，G418筛选出稳定表达株。并利用RT—PCR、免疫组织化学方法证明目的基因能够在骨髓基质干细胞中得到稳定表达。MTT实验结果显示转基因后的MSCs增殖能力有不同程度的提高。细胞内碱性磷酸酶活性明显上调，BMP-2、bFGF转染组骨钙索水平升高，成骨活性增强。提示：BMP-2、bFGF基因修饰的骨髓基质干细胞作为种子细胞能够更为有效地在骨组织工程中发挥作用。

The Study of Osteogenous Differentiation of MSCs Transfected with Different Gene

This experiment is sought to study the contribution of different gene in osteogenous differentiation of bone marrow stromal cells(MSCs) and optimize the seed cells of bone tissue engineering.Firstly,we obtained the full length gene of BMP-2,VEGF165 and bFGF by RT-PCR,and cloned into the expression vector pcDNA3.0.After to be transfected,the MSCs cell lines which could express each of the target protein were selected out by G418.RT-PCR and immunohistochemistry were used to confirm the exogenous gene expression in MSCs.MTT showed that almost all of the MSCs which have been transfected with exogenous gene had more strong proliferative potential than the untransfected group did.There was a notable increase of ALP activity in transfected cell compared with the control group.The concentration of OCN in cell culture medium had a increase in some degree except of VEGF group.The outstanding osteogenous differentiation could be observed in BMP-2 and bFGF transfected MSCs.The results showed that the MSCs modified by BMP-2 and bFGF would be more effective in bone tissue engineering field.