慢病毒介导GFP转染小鼠ESC及其对干细胞特性的影响

龙源期刊网 https://www.wendangku.net/doc/7b5274391.html,

慢病毒介导GFP转染小鼠ESC及其对干细胞特性的影响

作者：张爱霞

来源：《中国实用医药》2015年第11期

【摘要】目的研究用慢病毒载体进行基因转染对小鼠胚胎干细胞（ESC）特性的影响。方法慢病毒载体经293FT细胞包装后转染小鼠ESC，荧光显微镜下挑选和观察所得阳性克隆，检测ESC特异性标志物Oct-4、SSEA-1的表达，通过拟胚体（EB）诱导体外分化， RT-PCR检测三胚层的分化。结果形态学观察可见所得绿色荧光蛋白（GFP）阳性细胞呈克隆样

鸟巢状生长，免疫组化可见GFP-ESC表达ESC特异性标志Oct-4、SSEA-1，能形成EB，

RT-PCR检测到甲胎蛋白（AFP）、巢蛋白（Nestin）和血管内皮生长因子受体（Flk-1）的表达。结论慢病毒载体介导的基因转染不影响小鼠ESC的自我更新特性及多向分化潜能。

【关键词】胚胎干细胞；慢病毒载体；基因转染；绿色荧光蛋白

DOI：10.14163/https://www.wendangku.net/doc/7b5274391.html,ki.11-5547/r.2015.11.002

Lentiviral mediated GFP in transfection of mice ESC and its influence on stem cell features ZHANG Ai-xia. School of Bioscience， Jiaying College， Meizhou 514015， China

【Abstract】 Objective To study the influence of gene transfection by lentiviral vector on mice embryonic stem cell （ESC） features. Methods Transfection of mice ESC was made by lentiviral vector after 293FT cell packing. Positive clone was chosen and observed under fluorescence microscope. Expressions of Oct-4 and SSEA-1 as ESC specific markers were detected. In vitro differentiation was induced by embryoid body （EB）， and RT-PCR was applied to detect differentiation of triploblastica. Results Green fluorescent protein （GFP） positive cell showed colon bird-nest growth by morphological observation. Immunohistochemistry showed Oct-4 and SSEA-1 as ESC specific markers could form EB. Expressions of alpha fetoprotein （AFP）， nestin and vascular endothelial growth factor receptor （Flk-1） could be detected by RT-PCR. Conclusion Gene transfection mediated by lentiviral vector has no influence on self-renewal feature and multi-directional differentiation potency of mice ESC.

【Key words】 Embryonic stem cell； Lentiviral vector； Gene transfection； Green fluorescent protein

胚胎干细胞（embryonic stem cells， ESC）是从附植前早期胚胎内细胞团或早期胎儿原始生殖细胞经体外分离培养而来的多能性细胞系。ESC因能够无限增殖并保持未分化状态和具有向3个胚层细胞分化的潜力，在研究胚胎发育关键基因功能、建立疾病模型等方面有着其他

细胞无法比拟的优势。转基因研究是更有效利用ESC进行各种基因功能研究以及基因治疗等