借助慢病毒将EGFP基因导入西藏小型猪胎儿成纤维细胞  

Lentivirus-Mediated Gene Transfer of EGFP Gene into Porcine Embryonic Fibroblasts (PEFs) of Tibetan Miniature Pigs

作　　者：张晟[1] 肖高芳[2] 黄黎珍[1] 那顺巴雅尔[1] 贾俊双[2] 姚志芳[2] 肖东[1] 顾为望[1] 

机构地区：[1]南方医科大学比较医学研究所暨实验动物中心,广州510515 [2]南方医科大学肿瘤研究所,广州510515

出　　处：《中国比较医学杂志》2010年第4期34-36,55,88,共5页Chinese Journal of Comparative Medicine

基　　金：广东省自然科学基金项目(9151063101000015);广东省科技计划项目(2009B060300008);广东省医学科学技术研究基金(A2007359);南方医科大学优秀中青年科技人才库科研资助金;广州地区科学仪器协作共用网专用基金(2006176)

摘　　要：目的借助慢病毒将EGFP基因导入西藏小型猪胎儿成纤维细胞(porcine embryonic fibroblasts,PEFs),以基于PEFs建立慢病毒介导的外源基因体外投递系统。方法取35d西藏小型猪胚胎,酶消化法分离培养西藏小型猪的PEFs;按Invitrogen公司推荐的标准程序进行慢病毒(携带EGFP基因)包装(脂质体介导的瞬时转染),随后用病毒上清感染PEFs,24～48h后荧光显微镜下观察是否见绿色荧光以证实慢病毒是否成功生产和成功感染PEFs。结果成功分离培养西藏小型猪的PEFs,按标准程序生产的携带EGFP基因慢病毒高效率感染西藏小型猪的PEFs。结论针对西藏小型猪的PEFs建立了相应的慢病毒介导的外源基因体外投递系统,为相关后续研究打下了良好基础。Objective To establish lentivirus-mediated in vitro gene delivery systems for porcine embryonic fibroblasts（PEFs） of Tibetan miniature pigs.Methods According to the standard protocol from Invitrogen,lentiviruses carrying EGFP gene were produced,followed by confirming that lentiviruses harboring EGFP gene were successfully made and PEFs were successfully infected through EGFP assay under fluorescent stereo microscope after infecting PEFs.Results PEFs of Tibetan miniature pigs were successfully and efficiently infected by lentivirus supernatant produced according to the standard protocol from Invitrogen.Conclusions Lentivirus-mediated in vitro gene delivery systems for PEFs of Tibetan miniature pigs have been successfully established in our laboratory.

关 键 词：慢病毒 西藏小型猪胎儿成纤维细胞(PEFs) 绿色荧光蛋白 

分 类 号：R332[医药卫生—人体生理学]