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| 腺相关病毒介导色素上皮衍生因子转染人虹膜色素上皮细胞的体外研究 | |
| 引用本文： | 赵莼,王方,计菁,施沃栋,范先群,罗敏.腺相关病毒介导色素上皮衍生因子转染人虹膜色素上皮细胞的体外研究[J].上海交通大学学报(医学版),2011,31(8):1078-1081. |
| 作者姓名： | 赵莼 王方 计菁 施沃栋 范先群 罗敏 |
| 作者单位： | 1. 上海交通大学医学院附属第九人民医院眼科,上海 200011;同济大学医学院附属第十人民医院眼科,上海 200072 2. 同济大学医学院附属第十人民医院眼科,上海,200072 3. 上海交通大学医学院附属第九人民医院眼科,上海,200011 |
| 基金项目： | 上海市卫生局基金(2006028)~~ |
| 摘 要： | 目的探讨以腺相关病毒(AAV)为载体,对体外培养的人虹膜色素上皮(IPE)细胞进行色素上皮衍生因子(PEDF)转染,获得高效表达PEDF转基因细胞的可行性。方法构建携带PEDF基因的重组AAV载体AAV-PEDF,体外培养人IPE,用1×106pfu的AAV-PEDF按感染复数(MO I)为0、2、10和20分别感染体外培养的IPE细胞;荧光显微镜下观察细胞绿色荧光蛋白表达情况;流式细胞术检测细胞转染效率;RT-PCR和W estern b lotting法鉴定PEDF在IPE细胞中的表达。结果 AAV-PEDF转染后,IPE细胞生长正常,用荧光显微镜、RT-PCR和W estern b lotting均检测到PEDF表达。在MO I为20时,细胞感染阳性率达64.22%。结论 AAV-PEDF能有效地转染体外培养的IPE细胞并有效表达PEDF。 |
| 关 键 词： | 腺相关病毒 色素上皮衍生因子 虹膜色素上皮细胞 转染 |
Adeno-associated virus mediated transduction of pigment epithelium-derived factor to human iris pigment epithelial cells in vitro |
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| ZHAO Chun,WANG Fang,JI Jing,SHI Wo-dong,FAN Xian-qun,LUO Min.Adeno-associated virus mediated transduction of pigment epithelium-derived factor to human iris pigment epithelial cells in vitro[J].Journal of Shanghai Jiaotong University:Medical Science,2011,31(8):1078-1081. | |
| Authors: | ZHAO Chun WANG Fang JI Jing SHI Wo-dong FAN Xian-qun LUO Min |
| Affiliation: | ZHAO Chun1,2,WANG Fang2,JI Jing1,SHI Wo-dong1,FAN Xian-qun1,LUO Min1(1.Department of Ophthalmology,the Ninth People's Hospital,Shanghai Jiaotong University School of Medicine,Shanghai200011,China,2.Department of Ophthalmology,the Tenth People's Hospital,Tongji University,Shanghai200072,China) |
| Abstract: | Objective To investigate the feasibility of recombinant adeno-associated virus(AAV) mediated transduction of pigment epithelium-derived factor(PEDF) to cultured human iris pigment epithelial(IPE) cells in vitro to obtain the transgenic cells with high expression of PEDF. Methods Recombinant AAV vector carrying PEDF gene(AAV-PEDF) was constructed,and human IPE cells were cultured in vitro and infected with condensed virus.The expression of green fluorescent protein in cells was observed under fluorescence mi... |
| Keywords: | adeno-associated virus pigment epithelium-derived factor iris pigment epithelial cell transduction |
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