增强型绿色荧光蛋白转染猪骨髓间充质干细胞特性观察  

Observation of Biological Characteristics of Porcine Mesenchymal Stem Cells after Enhance Green Fluorescent Protein Gene Transfer

作　　者：牛德亮[1] 常彩红[2] 

机构地区：[1]河南中医学院第一附属医院输血科,河南郑州450000 [2]郑州大学第三附属医院检验科,河南郑州450052

出　　处：《中国误诊学杂志》2009年第23期5551-5553,共3页Chinese Journal of Misdiagnostics

基　　金：河南省教育厅自然基金资助项目(编号:2007310014)

摘　　要：目的:借助质粒将增强型绿色荧光蛋白(EGFP)基因转染猪骨髓干细胞(MSC),观察转染后EGFP在(MSC)内表达情况及MSC生长特性。方法:1月龄小型香猪为MSC供体,EGFP基因转染MSC,计算转染效率,流式细胞仪鉴定G418筛选后EGFP基因稳定表达率和转染后细胞增殖周期改变。结果:转染率为(36.9±3.4)%;G418筛选后EGFP表达率75.6%,EGFP转染MSC G0/G1期MSC比例较对照组明显增加(P<0.01),S+G2/M期MSC比例减少(P<0.01)。结论:猪MSC经基因转染和G418筛选,获得EGFP稳定表达;转染和G418筛选后MSC增殖能力降低,G0/G1期MSC比例增加,S+G2/M期MSC比例下降。Objective：To transfer EGFP gene into porcine MSCs by adopting IRE-S2, EGFP vector so as to observe the expression of enhance green fluorescent protein （EGFP） and their growth characters after gene transfer, to provide the evidence for MSCs existing and fates in the future research. Methods：Chinese Laboratory Miniature pigs aged 1 months were used as donors of bone marrow stem cells. We transfered EGFP gene into porcine MSCs. The expression of EGFP was observed and the ratio of transfection was calculated. The expression ratio of EGFP gene and the proliferation ability was detected using FACS after G418 selecting. Results：EGFP expression ratio was 36.9%. EGFP expression was up to 75.6% after G418 selecting. Compared with untransfected control MSCs, the number of cells increased in G0/G1 phase （P〈0. 01）and decreased in ,S＋G2/M phase（P〈0. 01）. Conclusion ： Steady expression of EGFP is in MSCs by the gene transfer technique and G418 selecting. The proliferate activity is inhibited after gene transfer. The number of cells increase in G0/G1 phase and decrease in S＋G2/M phase.

关 键 词：绿色荧光蛋白质类 猪/遗传学 间质干细胞/细胞学 动物 

分 类 号：R394.2[医药卫生—医学遗传学]