借助慢病毒将EGFP基因导入西藏小型猪胎儿成纤维细胞

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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35天西藏小型猪胚胎,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。
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