Lentivirus vector-mediated enhanced green fluorescence protein transfection on in vitro corneal epithelial cells of rabbits

王继东; 杜之渝; 兰芬

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Lentivirus vector-mediated enhanced green fluorescence protein transfection on in vitro corneal epithelial cells of rabbits

更新时间：2026-01-09

- Lentivirus vector-mediated enhanced green fluorescence protein transfection on in vitro corneal epithelial cells of rabbits
- Journal of Army Medical University Vol. 32, Issue 13, Pages: 1425-1428(2010)
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  CLC： R322.91;R394-33;R394.2
- Published：2010
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王继东,杜之渝,兰芬.慢病毒载体介导增强型绿色荧光蛋白转导兔角膜上皮细胞的体外实验研究[J].陆军军医大学学报(原第三军医大学学报),2010,32(13):1425-1428.

　Wang Jidong,Du Zhiyu,Lan Fen.Lentivirus vector-mediated enhanced green fluorescence protein transfection on in vitro corneal epithelial cells of rabbits[J].J Amry Med Univ (J Third Mil Med Univ),2010,32(13):1425-1428.
王继东,杜之渝,兰芬.慢病毒载体介导增强型绿色荧光蛋白转导兔角膜上皮细胞的体外实验研究[J].陆军军医大学学报(原第三军医大学学报),2010,32(13):1425-1428. DOI：

　Wang Jidong,Du Zhiyu,Lan Fen.Lentivirus vector-mediated enhanced green fluorescence protein transfection on in vitro corneal epithelial cells of rabbits[J].J Amry Med Univ (J Third Mil Med Univ),2010,32(13):1425-1428. DOI：

摘要

目的观察慢病毒载体（lentiviral vectors

LVs）用于兔角膜上皮细胞基因转染的有效性。方法兔角膜上皮细胞的原代及传代培养并做细胞鉴定；慢病毒载体介导的增强型绿色荧光蛋白（lenti-EGFP）以不同的感染复数（multiplicity of infection

MOI=0、1、10、50、100、500）转染实验细胞，寻找最佳转染剂量；于转染后24、48、72、96 h运用倒置荧光显微镜观察不同MOI下增强型绿色荧光蛋白（enhanced green fluorescence protein

EGFP）的表达并计算细胞转染率；RT-PCR 方法检测EGFP基因的表达情况。结果 EGFP于转染48 h即开始有表达，随着转染时间的延长其表达增强。MOI=1、10、50、100时，角膜上皮细胞转染率随着感染复数的增加而增加，分别为(4.5±0.6）%、（30.4±0.9）%、（51.9±1.4）%、（75.4±0.7）%，各组间差异显著（P0.05）；MOI在100与500时，转染率差异不显著（P0.05），即最适感染复数为100。RT-PCR结果提示转染细胞组EGFP基因有表达。结论慢病毒载体能够有效转染离体兔角膜上皮细胞。

Abstract

Objective To observe the effect of lentivirus vector-medicated enhanced green fluorescence protein (EGFP) transfection on in vitro corneal epithelial cells. Methods Primary corneal epithelial cells of rabbits were cultured and transfected with lentivirus vector into different multiplicity of infection (MOI) in order to find the best transfection dose. After 24

and 96 h of transfection

expression of EGFP was observed under microscope and detected by RT-PCR. The transfection rate of corneal epithelial cells was calculated in different MOI. Results EGFP was expressed in corneal epithelial cells 48 h after transfection

which increased with the increasing transfection time. The transfection rate of corneal epithelial cells was 4.5%±0.6%

30.4%±0.9%

51.9%±1.4%

and 75.4%±0.7%

respectively

when MOI was 1

and 100（P0.05). However

no difference was found in the transfection rate of corneal epithelial cells when MOI was 100 and 500. The optimol MOI was 100. RT-PCR showed that EGFP gene was expressed in transfected corneal epithelial cells. Conclusion Lentivirus vector can effectively transfect corneal epithelial cells in vitro.

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