• 主题
高级搜索  >
历史搜索 清空历史
首页> 中文期刊> 《实验动物与比较医学》 >西藏小型猪骨髓间充质干细胞分离培养及慢病毒载体介导EGFP标记间充质干细胞

西藏小型猪骨髓间充质干细胞分离培养及慢病毒载体介导EGFP标记间充质干细胞

         
页面导航
  • 摘要
  • 著录项
  • 相似文献
  • 相关主题

摘要

Objective To establish a simple and effective method used to isolate and purify bone marrow mesenchymal stem cells (BMSCs) of Tibetan miniature pigs, and further set up the lentivirus-mediated in vitro gene delivery system for BMSCs of Tibetan miniature pigs. Methods ① MSCs of Tibetan miniature pigs were initially isolated by density gradient centrifugation, followed by purifying MSCs using differential attachment method, and further passage 3 BMSCs of Tibetan miniature pigs were characterized by CD45, CD90 and CD 105.② According to the standard protocol from Invitrogen, lentiviruses carrying EGFP gene were produced, and then infected MSCs, followed by EGFP assay under fluorescent stereo microscope and fluorescence activated cell sorting (FACS) for determining the infecting efficiency. Results BMSCs of Tibetan miniature pigs obtained by the combined use of density gradient centrifugation and differential attachment method were shown to enjoy the characteristics of MSCs. Additionally, the BMSCs of Tibetan miniature pigs were successfully and efficiently infected by lentivirus supernatant, and the infecting efficiency was 84.1%. Conclusion High purified BMSCs of Tibetan miniature pigs can be obtained by the combined use of density gradient centrifugation and differential attachment method, and lentivirus-mediated in vitro gene delivery systems for BMSCs of Tibetan miniature pigs were successfully established.%目的 建立一种简便、有效的西藏小型猪骨髓间充质干细胞(BMSCs)体外分离培养及纯化方法,并基于BMSCs建立慢病毒介导的外源基因EGFP体外投递系统.方法 密度梯度离心法分离西藏小型猪BMSCs,差速贴壁法不断纯化,流式细胞仪检测第3代BMSCs CD45、CD90和CD105表达；此外,用携带EGFP基因的慢病毒转染BMSCs以获得EGFP标记的BMSCs,倒置荧光显微镜下观察感染情况及流式细胞术检测感染效率.结果 ①密度梯度分离法结合差速贴壁法所获得的细胞经鉴定符合MSCs的特性.②借助慢病毒高效率将EGFP导入BMSCs,感染效率为84.1％.结论 利用密度梯度分离法结合差速贴壁法可获得高纯度的BMSCs,基于BMSCs 建立了慢病毒介导的外源基因体外投递系统.

著录项

相似文献

  • 中文文献
  • 外文文献
  • 专利
获取原文

客服邮箱：kefu@zhangqiaokeyan.com

京公网安备：11010802029741号 ICP备案号：京ICP备15016152号-6 六维联合信息科技 (北京) 有限公司©版权所有

  • 客服微信

  • 服务号