Tai Dapeng, Liu Pengxia, Gao Jing, Jin Muzi, Xu Teng, Zuo Yongchun, Liang Hao, Liu Dongjun
1 Key Laboratory of Mammalian Reproductive Biology and Biotechnology Ministry of Education, Inner Mongolia University , Inner Mongolia, Hohhot, 010021, China .
2 These authors contributed equally to this work.
Cell Reprogram. 2015 Aug;17(4):297-305. doi: 10.1089/cell.2014.0107.
Various factors affect the process of obtaining stable Arbas cashmere goat embryonic stem cells (ESCs), for example, the difficulty in isolating cells at the appropriate stage of embryonic development, the in vitro culture environment, and passage methods. With the emergence of induced pluripotent stem cell (iPSC) technology, it has become possible to use specific genes to induce somatic cell differentiation in PSCs. We transferred OCT4, SOX2, c-MYC, and KLF4 into Arbas cashmere goat fetal fibroblasts, then induced and cultured them using a drug-inducible system to obtain Arbas goat iPSCs that morphologically resembled mouse iPSCs. After identification, the obtained goat iPSCs expressed ESC markers, had a normal karyotype, could differentiate into embryoid bodies in vitro, and could differentiate into three germ layer cell types and form teratomas in vivo. We used microarray gene expression profile analysis to elucidate the reprogramming process. Our results provide the experimental basis for establishing cashmere goat iPSC lines and for future in-depth studies on molecular mechanism of cashmere goat somatic cell reprogramming.
多种因素影响着获取稳定的阿尔巴斯绒山羊胚胎干细胞(ESCs)的过程,例如,在胚胎发育的合适阶段分离细胞的困难程度、体外培养环境以及传代方法。随着诱导多能干细胞(iPSC)技术的出现,利用特定基因诱导体细胞分化为多能干细胞已成为可能。我们将OCT4、SOX2、c-MYC和KLF4导入阿尔巴斯绒山羊胎儿成纤维细胞,然后使用药物诱导系统对其进行诱导培养,以获得形态上与小鼠iPSCs相似的阿尔巴斯山羊iPSCs。经鉴定,所获得的山羊iPSCs表达胚胎干细胞标志物,具有正常的核型,能够在体外分化为胚状体,并且能够在体内分化为三种胚层细胞类型并形成畸胎瘤。我们使用基因芯片基因表达谱分析来阐明重编程过程。我们的结果为建立绒山羊iPSC系以及未来深入研究绒山羊体细胞重编程的分子机制提供了实验依据。