mdr1基因重组腺病毒转染小鼠单个核细胞对其胞内结构的影响

Influence of mice mononuclear cells transfected with Ad-EGFP-mdr1 on ultrastructure

目的用表达EGFP和mdr1基因重组腺病毒转染原代培养的小鼠单个核细胞,研究该重组腺病毒在其细胞表达的安全性。方法将重组腺病毒Ad-EGFP-mdr1转染原代培养的Babl/c小鼠单个核细胞,以EGFP做报告基因用荧光倒置显微镜观察其在细胞内的表达。用透射电镜观察其在胞内分布。结果荧光显微镜下可观察到转染成功的原代培养小鼠单个核细胞发绿色荧光。电镜检查其胞内有大量病毒颗粒存在,细胞超微结构正常。结论用EGFP检测转染重组腺病毒Ad-EGFP-mdr1的单个核细胞方法简单,其线粒体、内质网、细胞核等超微结构无病理性改变。实验证明转染是安全有效的。为进一步研究转染Ad-EGFP-mdr1单个核细胞回输的骨髓保护作用的安全性奠定基础。

Objective To observe the influence of mice mononuclear cells transfected with recombinant adenovirus containing EGFP and mdrl on ultrastructure. Methods Mice mononuclear cells were primarily cultured and transfected with Ad-EGFP-mdr1. EGFP,the reporter gene, was observed by fluorescence inverted microscope. The ultrastructures and the viruses were located by transmission electron microscope. Results mdrl represented by EGFP was expressed in the experiment group. Many of viruses were in the mononuclear cells and the ultrastructures were not abnomal. Conclusion It is safe to transfect the Ad-EGFP-mdr1 into mice mononuclear cells.