上海交通大学学报（医学版）

›› 2013, Vol. 33 ›› Issue (4): 439-.doi: 10.3969/j.issn.1674-8115.2013.04.012

• 论著（基础研究） • 上一篇    下一篇

慢病毒负载GFP转染兔脂肪干细胞的效率及毒性评价

李宏松1, 徐 晨2,3, 邹 俊1   

  1. 1.上海交通大学附属第六人民医院眼科, 上海 200233； 2.上海交通大学 基础医学院组织胚胎学教研室, 上海 200025； 3.上海市生殖医学重点实验室, 上海 200025
  • 出版日期:2013-04-28 发布日期:2013-05-03
  • 通讯作者: 邹 俊, 电子信箱: zoujun70@126.com。
  • 作者简介:李宏松（1987—）, 男, 硕士生； 电子信箱: lihongsong1987@126.com。

Transfection efficiency and cytotoxicity of lentivirus-GFP on rabbit adipose-derived stem cells

LI Hong-song1, XU Chen2,3, ZOU Jun1   

  1. 1.Department of Ophthalmology, the Sixth People´s Hospital, Shanghai Jiaotong University, Shanghai 200233, China； 2.Department of Histology and Embryology, Basic Medical College, Shanghai Jiaotong University, Shanghai 200025, China； 3.Shanghai Key Laboratory of Reproductive Medicine, Shanghai 200025, China
  • Online:2013-04-28 Published:2013-05-03

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摘要：

目的 探讨慢病毒负载绿色荧光蛋白（GFP）对兔脂肪干细胞(rADSCs)转染的可行性及稳定性，为rADSCs组织工程化角膜上皮的研究提供可靠的体内外示踪方法。方法 以慢病毒作为载体，采用不同感染复数（MOI）慢病毒将GFP基因导入rADSCs，荧光倒置显微镜下观察细胞转染情况，应用MTT法检测慢病毒转染对rADSCs细胞生长的影响，评价细胞毒性。结果 当MOI值为50、100、500时，rADSCs均可被成功转染，120 h后转染效率均可达50%以上。当MOI值为50、100时，GFP-慢病毒对rADSCs的生长无明显影响，而MOI值为500时，细胞生长受到明显抑制。结论 慢病毒负载GFP可成功导入rADSCs，为进一步对ADSCs组织工程化角膜上皮的研究奠定了良好的实验基础。

关键词: 脂肪干细胞, 毒性评价, 绿色荧光蛋白, 转染, 兔

Abstract:

Objective To evaluate the transfection efficiency and cytotoxicity of lentivirus containing green fluorescence protein (GFP)(lentivirus-GFP) on rabbit adipose-derived stem cells (rADSCs), and establish a method of cell tracer marker in the study of reconstruction of tissue engineered corneal epithelium. Methods rADSCs were transfected with lentivirus-GFP at different multiplicity of infection (MOI). Cell transfection was observed under fluorescent microscope, and influence of lentivirus transfection on proliferation of rADSCs was detected by MTT. Results rADSCs were successfully transfected at MOI of 50, 100 and 500, and the transfection efficiency could reach more than 50% 120 h later. Lentivirus-GFP had no significant effect on proliferation of rADSCs at MOI of 50 and 100, while the proliferation of rADSCs was significantly inhibited at MOI of 500. Conclusion rADSCs are successfully transfected with lentivirus-GFP, which can be used in the study of reconstruction of tissue engineered corneal epithelium.

Key words: adipose-derived stem cells, toxicity evaluation, green fluorescent protein, transfection, rabbit