[1]郑钧丰,孔永,葛良鹏,等.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].第三军医大学学报,2008,30(12):1111-1113.
 ZHENG Jun-feng,KONG Yong,GE Liang-peng,et al.AAV2-mediated versus AAV2/1-mediated enhanced green fluorescent protein transfection into human skin fibroblast in vitro[J].J Third Mil Med Univ,2008,30(12):1111-1113.
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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(/HTML )
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《第三军医大学学报》[ISSN:1000-5404/CN:51-1095/R]

卷:
30卷
期数:
2008年第12期
页码:
1111-1113
栏目:
论著
出版日期:
2008-06-30

文章信息/Info

Title:
AAV2-mediated versus AAV2/1-mediated enhanced green fluorescent protein transfection into human skin fibroblast in vitro
作者:
郑钧丰孔永葛良鹏魏泓
第三军医大学基础医学部实验动物学教研室; 西南大学生命科学院; 重庆宗申医达公司
Author(s):
ZHENG Jun-feng KONG Yong GE Liang-peng WEI Hong
Department of Laboratory Animal Science, College of Medicine, Third Military Medical University, Chongqing 400038; School of Life Science, Southwest University, Chongqing 400017;Zong Shen Yida Company, Chongqing 400054, China
关键词:
腺相关病毒人皮肤成纤维细胞增强型绿色荧光蛋白流式细胞
Keywords:
adeno-associated virus human fibroblast enhanced green fluorescent protein flow cytometry
分类号:
R329;R373;R394-33
文献标志码:
A
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104、105、106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104时转染效率为(7.68±1.18)%,当MOI达到105后,转染效率稳定在(22.16±5.59)%,随着MOI增加,最高至MOI 106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106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 104 to 106. 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 104, the transfection efficiency of AAV2-EGFP was 7.68%±1.18%; When MOI was 105, that was 52.12%±1.59%; When MOI was 106, 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.

参考文献/References:

郑钧丰,孔永,葛良鹏,等.  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第三军医大学学报,2008,30(12):1111-1113.

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更新日期/Last Update: 2008-06-13