AAV2-mediated versus AAV2/1-mediated enhanced green fluorescent protein transfection into human skin fibroblast in vitro

郑钧丰; 孔永; 葛良鹏; 魏泓

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AAV2-mediated versus AAV2/1-mediated enhanced green fluorescent protein transfection into human skin fibroblast in vitro

更新时间：2026-01-09

- AAV2-mediated versus AAV2/1-mediated enhanced green fluorescent protein transfection into human skin fibroblast in vitro
- Journal of Army Medical University Vol. 30, Issue 12, Pages: 1111-1113(2008)
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  CLC： R329;R373;R394-33
- Published：2008
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郑钧丰,孔永,葛良鹏,魏泓.2种腺相关病毒介导的增强型绿色荧光蛋白对人成纤维细胞转染效率的研究[J].陆军军医大学学报(原第三军医大学学报),2008,30(12):1111-1113.

　ZHENG Jun-feng,KONG Yong,GE Liang-peng,WEI Hong.AAV2-mediated versus AAV2/1-mediated enhanced green fluorescent protein transfection into human skin fibroblast in vitro[J].J Amry Med Univ (J Third Mil Med Univ),2008,30(12):1111-1113.
郑钧丰,孔永,葛良鹏,魏泓.2种腺相关病毒介导的增强型绿色荧光蛋白对人成纤维细胞转染效率的研究[J].陆军军医大学学报(原第三军医大学学报),2008,30(12):1111-1113. DOI：

　ZHENG Jun-feng,KONG Yong,GE Liang-peng,WEI Hong.AAV2-mediated versus AAV2/1-mediated enhanced green fluorescent protein transfection into human skin fibroblast in vitro[J].J Amry Med Univ (J Third Mil Med Univ),2008,30(12):1111-1113. DOI：

摘要

目的比较2种不同腺相关病毒(adeno-associated virus

AAV)介导的增强型绿色荧光蛋白对人皮肤成纤维细胞（human fibroblast

HFB）的转染效率。方法原代分离培养人皮肤成纤维细胞并鉴定，按照感染复数（multiple of infection，MOI）为10

、10

转染传2代的人成纤维细胞，流式细胞检测AAV2-EGFP和AAV2/1-EGFP对HFB的转染效率

倒置荧光显微镜观察转染后的荧光强度。MTT法检测AAV2-EGFP和AAV2/1-EGFP对HFB的毒性。结果腺相关病毒AAV2EGFP对HFB的转染效率随着MOI增加而增高

MOI为10

时转染效率为(7.68±1.18)%，当MOI达到10

后，转染效率稳定在(22.16±5.59)%

随着MOI增加，最高至MOI 10

转染效率无显著提高。而病毒AAV2/1对于HFB的转染效率在MOI 为10

转染效率仅为(3.47±0.93)%。2种病毒在转染过程中，细胞生长正常

MTT显示各转染组与未转染组的吸光度值无统计学差异。结论 AAV2载体对体外培养的HFB转染效率高于AAV2/1，但2种病毒载体对人皮肤成纤维细胞转染效率均不高，AAV2/1-EGFP对HFB几乎无感染能力。

Abstract

Objective To compare the transfection rate of AAV2-EGFP and AAV2/1-EGFP to human skin fibroblast (HFB). Methods Two recombinant adeno-associated viruses (AAV) encoding enhanced green fluorescent protein (EGFP) were constructed and transfected to HFB at multiple of infection MOI ranging from 10

to 10

. Twenty-four hours after the infection

the expression rates of

EGFP on cultured HFB were assessed by flow cytometry and the transfected cells were observed under fluorescence microscope. The killing effect of the virus on the infected cells was assayed by MTT. Results The transfection efficiency of AAV2-EGFP was increased as MOI increased. When MOI was 10

the transfection efficiency of AAV2-EGFP was 7.68%±1.18%; When MOI was 10

that was 52.12%±1.59%; When MOI was 10

that showed no significant increase. However

the transfection efficiency of AAV2/1-EGFP showed no obvious changes at any MOI. Conclusion AAV2 is more efficient than AAV2/1 in transfecting HFB

but neither AAV has a high transfection efficiency.

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