限定因子诱导胎猪成纤维细胞重编程为多能性细胞

Generation of Induced Pluripotent Stem Cells From Porcine Fibroblasts

尝试运用限定因子融合蛋白建立猪的诱导多能性干细胞.试验采用Oct4、Sox2、Klf4、c-Myc四种限定因子经慢病毒表达载体系统介导感染猪胎儿成纤维细胞,对表达外源限定因子的猪胎儿成纤维细胞进行培养传代,逐步分离培养出集落边缘界限清晰的细胞克隆,细胞集落生长状态稳定、核型正常、碱性磷酸酶检测为阳性,免疫细胞化学检测显示,Oct4、Nanog、SSEA-1蛋白表达为阳性,体内能够分化形成含有三个胚层的畸胎瘤.结果证实分离培养的细胞克隆为猪诱导多能性干细胞,为进一步完善诱导方案和深入研究应用猪诱导多能性干细胞奠定了基础.

In order to establish pig induced pluripotent stem cells （iPS） with defined factor fusion protein, four defined factors genes Oct4, Sox2, c-Myc and Klf4 were delivered into porcine fetal fibroblasts by lentiviral transfection. The porcine fetal fibroblasts expressed exogenous defined factor genes were sub-cultured, and the clear-cut cell clones were gradually isolated. The cell colones grew at similar rates and stability, exhibited normal karyotype, and expressed alkaline phosphatase, Oct4, Nanog and SSEA1. And these cells could differentiate into various kinds of tissue in teratomas. The results confirmed that the isolated cell clones were iPS cells. This would greatly facilitate the further improvement of the induction protocol and in-depth study and application of pig iPS cells.