上海交通大学学报(医学版)

• 技术与方法 • 上一篇    

动物活体成像生物发光间充质干细胞系的构建

陈洁玉,施小凤   

  1. 江苏大学附属医院血液科, 镇江 212001
  • 出版日期:2014-11-28 发布日期:2014-12-02
  • 通讯作者: 施小凤, 电子信箱: shixiaofeng1977@163.com。
  • 作者简介:陈洁玉(1977—), 女, 主治技师, 学士; 电子信箱: shengsjm@sohu.com。
  • 基金资助:

    镇江市科技计划项目(SH2012029);中国博士后科学基金项目(2013M541528)

Establishment of mesenchymal stem cell lines with optical in vivo imaging ability and bioluminescence

CHEN Jie-yu, SHI Xiao-feng   

  1. Department of Hematology, Affiliated Hospital of Jiangsu University, Zhenjiang 212001, China
  • Online:2014-11-28 Published:2014-12-02
  • Supported by:

    Project of Scienceand Technology Commission of Zhenjiang Municipality, SH2012029; China Postdoctoral Science Foundation, 2013M541528

摘要:

目的 建立稳定表达荧光素酶的小鼠骨髓间充质干细胞(MSCs)系。方法 用携带荧光素酶基因的慢病毒GV260感染F3代MSCs系OP9细胞,通过嘌呤霉素筛选建立稳定表达荧光素酶的OP9luc+细胞;检测荧光素酶报告基因的活性,观察其表达效果;将OP9luc+细胞经尾静脉注射至C57BL小鼠体内,观察其成像能力。结果 筛选到稳定表达荧光素酶的小鼠MSCs系OP9luc+;活性测定证明其表达效率高;在动物体内可以清晰成像。结论 构建了稳定高表达荧光素酶基因的小鼠MSCs系OP9luc+。

关键词: 荧光素酶, 活体成像, 生物发光, 骨髓间充质干细胞

Abstract:

Objective To establish murine bone marrow mesenchymal stem cell lines that can stably express the luciferase. Methods OP9 cells of MSC lines were infected by lentivirus GV260 containing luciferase gene and screened by puromycin for establishing OP9 luc+ cell lines that could stably express the luciferase. The activity of reporter gene of luciferase was detected and its expression effect was observed. OP9 luc+ cells were injected via the tail vein of C57BL mice and imaging ability was observed. Results OP9 luc+ cells that stably expressed luciferase were screened. The activity detection proved that the expression efficiency of OP9 luc+ cells was high. OP9 luc+ cells could form image clearly in vivo. Conclusion Murine bone marrow mesenchymal stem cell lines with stable and high expression of luciferase are established.

Key words: luciferase, optical in vivo imaging, bioluminescence, bone marrow mesenchymal stem cell