腺相关病毒及慢病毒载体对骨髓间充质干细胞基因转染效率的比较  

Efficiency of Gene Transfection with Adeno- associated Viral Vector or Lentiviral Vector in Bone Marrow Derived Mesenchymal Stem Cells

作　　者：苏玉金[1] 赵育梅[1] 顾漪[1] 

机构地区：[1]北京市神经外科研究所首都医科大学附属北京天坛医院,北京市100050

出　　处：《中国康复理论与实践》2014年第12期1117-1121,共5页Chinese Journal of Rehabilitation Theory and Practice

基　　金：国家重大新药创制科技重大专项资助项目(No.2012ZX09401004)

摘　　要：目的比较腺相关病毒(AAV)载体和慢病毒(LV)载体在间充质干细胞(MSCs)基因转染中的效率。方法密度梯度离心法分离培养MSCs,HE染色、Nestin免疫荧光染色鉴定,Brd U标记观察增殖情况。分别包装AAV与LV假病毒颗粒,并感染MSCs,通过β-gal染色和绿色荧光蛋白检测,分别计算两者的转染效率。结果 MSCs成功从骨髓分离。AAV载体介导的基因转染率为49.1%,LV载体的转染率为91.4%(P<0.01)。结论 LV载体介导的基因转染方法转染效率更高。Objective To compare 2 kinds of commonly used viral vectors, adeno-associated viral（AAV） vector and lentiviral（LV） vector in the gene transfection for bone marrow derived mesenchymal stem cells（MSCs）. Methods MSCs were isolated with density gradient（lymphocytes seperation） and identified with HE staining and immunocytochemistory staining for Nestin. The proliferation of BMSCs was detected with Brd U labeling. AAV mediated gene transfection was carried out through recombinant AAV-Lac Z viral particles. For LV mediated gene transfection, the LV particles were used directly. The transfection efficiency was estimated with β-gal staining and green fluorescent protein under the fluorescent microscope respectively. Results MSCs was successfully isolated from the bone marrow. HE staining showed that MSCs was with big nucleus, 1-3 nucleoli, and high nucleocytoplasmic ratio. Brd U labeling suggested that MSCs were proliferating. Some MSCs expressed Nestin. The gene transfection efficiency mediated with AAV vector was 49.1%, and it was 91.4% with LV vector（P〈0.01）. Conclusion The LV vector is more efficient on gene transfection than AAV vector.

关 键 词：基因转染 腺相关病毒载体 慢病毒载体 骨髓间充质干细胞 

分 类 号：R394.8[医药卫生—医学遗传学]