端粒酶催化亚单位基因诱导人骨髓间充质干细胞的实验研究  

Immortalization of human bone marrow mesenchymal stem cells by transfected with hTERT

作　　者：刘新生[1] 许予明[2] 

机构地区：[1]郑州市第一人民医院神经内科,河南郑州450004 [2]郑州大学第一附属医院神经内科,河南郑州450052

出　　处：《中风与神经疾病杂志》2015年第6期532-534,共3页Journal of Apoplexy and Nervous Diseases

摘　　要：目的利用端粒酶催化亚单位基因(h TERT)异位表达建立永生化人骨髓间充质干细胞(h MSCs)系。方法使用含有h TERT的重组质粒载体转染人原代骨髓间充质干细胞,经新霉素G418筛选,连续传代培养,通过形态学、免疫组织化学及PCR技术等方法对细胞系进行鉴定。结果转染后骨髓间充质干细胞基本保持了原代细胞的表型特征、形态均一、多为梭形,呈漩涡状或放射状排列,有特征性表型。细胞免疫组化染色显示有h TERT的表达。结论成功地构建了h TERT永生化的人骨髓间充质干细胞系,为其在神经系统疾病治疗中的应用奠定了实验基础。Objective To immortalize human bone marrow mesenchymal stem cells（ h MSCs） by ectopic expression of human telomerase reverse transcriptase（ h TERT） without malignant transformation. Methods A vector that contained h TERT fragment and drug resistance gene were was used to transfect normal human bone marrow mesenchymal stem cells.The transfected cells were screened with 400 μg / ml G418. An under inverted microscope was used to observe the morphology and growth of the cells. The expressions of transfected DNA fragments were detected in expanded cell lines by immunocytochemistry and PCR. Results The cells were homogenous,closely apposed,large,flat,and polygonal,displayed a characteristic ovoid nucleus with one or two nucleoli and formed monolayer with polygonal shape without overlapping. The existence and expression of h TERT was detected in the transfected cells by immunocytochemistry and PCR. Immunochemistry results showed that the cultured cells were positive for CD29,CD44 and negative for CD34,CD45. Conclusion Ectopic expression of h TERT can effectively immortalize human bone marrow mesenchymal stem cells. It may provide stable cell resource for the basic researches and cell-transplantation therapies for basic and clinical research in neuroscience.

关 键 词：骨髓间充质干细胞 永生化 端粒酶催化亚单位基因 

分 类 号：R743[医药卫生—神经病学与精神病学]