目的 探索含Oct4、Sox2、C-Myc、Klf4四个基因的腺病毒载体感染小鼠的胚胎成纤维细胞技术,为制备诱导多能干细胞(iPSCs)研究奠定基础.方法 将胎鼠躯干用胰蛋白酶消化法分离培养小鼠胚胎成纤维细胞(MEF),将小鼠的胚胎成纤维细胞随机分为3组:A组:Oct4、Sox2、C-Myc、Klf4腺病毒载体感染组;B组:空病毒颗粒感染组;C组:正常对照组.用目的基因感染小鼠胚胎成纤维细胞,并观察其形态变化.结果 腺病毒感染小鼠胚胎成纤维细胞(MEF)的最佳感染复数(MOI)为20-100.A组细胞变为半椭圆形,B组和C组为为长梭形.结论 含Oct4、Sox2、C-Myc、Klf4四因子腺病毒载体成功感染了小鼠胚胎成纤维细胞,表明目的基因明显影响了MEF的形态,MEF细胞由长梭形变成了椭圆形或不规则形,为下一步实验奠定了基础.%Objective To explore the technology of adenovirus vector containingOct4,Sox2,C-Myc,Klf4four genesto infect mouse embryonic fibroblasts,and lay the foundation for the research on the preparation of induced pluripotent stem cells (iPSCs).Methods The mouse embryonic fibroblasts (MEF) were isolated and cultured by trypsin digestion method.The mouse embryonic fibroblasts were randomly divided into 3 groups:group A:Oct4,Sox2,C-Myc,Klf4 adenovirus vector infection group;group B:empty virus particle infection group;group C:normal control group.The mouse embryonic fibroblasts were infected with the target gene and the morphological changes were observed.Results The optimal number of infective complex (MOI) of adenovirus infected mouse embryonic fibroblasts (MEF) was 20-100.In group A,the cells became semi oval,and the cells of group B and group C were long spindle shape.Conclusion Mouse embryonic fibroblastswere successfully infected by adenovirus vector containing Oct4,Sox2,C-Myc,Klf4 four factors,indicating that the target gene significantly affected the morphology of MEF.MEF cells from long spindle shaped into an oval or irregular shape,laid the foundation for the next experiment.
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