pcDNA3-EGFP基因转染骨髓间充质干细胞的实验研究

Experimental study on pcDNA3-EGFP gene transfer to bone marrow mesenchymal stem cells

目的 探讨外源性pcDNA3-EGFP基因转染修饰兔骨髓间充质干细胞的可行性。方法 用增荧光绿色蛋白 (enhancedgreenfluorescentprotein ,EGFP)cDNA作为报告基因 ,采用重组真核表达载体系统 (pcDNA3-EGFP)以脂质体法转染原代骨髓间充质干细胞 ,观察转染结果、表达情况及对靶细胞活力的影响。结果 经荧光显微镜下观察证实 :成功地对骨髓间充质干细胞实现了基因转染 ,持续表达时间超过 8周 ;没有发现明显的细胞毒作用及对细胞活力的显著影响。结论 采用真核转染技术可以介导外源基因转染骨髓间充质干细胞 ,为今后采用基因增强组织工程方法治疗肌肉、骨骼系统疾病提供了实验基础。

Objective To study the feasibility of exogenous gene transfection to bone marrow mesenchymal stem cells (MSCs). Methods MSCs obtained from bone marrow of 4 adult Newzealand rabbits were cultured and treated with recombinant pcDNA3.1(+)-EGFP with the help of lipofectamine. The transgene results were confirmed by fluoroscopy and the cell viability post transfection was evaluated by light microscopy and electron microscopy. Results Bone marrow MSCs were successfully transfected with pcDNA3 carrying EGFP gene and the exogenous gene expression continued for over 8 weeks. Cell viability was not significantly influenced and no cell toxicity was observed. Conclusion Bone marrow MSCs can be efficiently modified by appropriate gene transfection through recombinant technique.