Jiangsu Engineering Technology Research Center of Meat Sheep & Goat Industry, Nanjing Agricultural University, Nanjing, People's Republic of China; Jiangsu Livestock Embryo Engineering Laboratory, Nanjing Agricultural University, Nanjing, People's Republic of China.
Mol Reprod Dev. 2013 Dec;80(12):1009-17. doi: 10.1002/mrd.22266. Epub 2013 Nov 27.
Embryonic stem cells (ESCs) are a powerful model for genetic engineering, studying developmental biology, and modeling disease. To date, ESCs have been established from the mouse (Evans and Kaufman, 1981, Nature 292:154-156), non-human primates (Thomson et al., , Proc Nat Acad Sci USA 92:7844-7848), humans (Thomson et al., 1998, Science 282:1145-1147), and rats (Buehr et al., , Cell 135:1287-1298); however, the derivation of ESCs from domesticated ungulates such as goats, sheep, cattle, and pigs have not been successful. Alternatively, induced pluripotent stem cells (iPSCs) can be generated by reprogramming somatic cells with several combinations of genes encoding transcription factors (OCT3/4, SOX2, KLF4, cMYC, LIN28, and NANOG). To date, iPSCs have been isolated from various species, but only limited information is available regarding goat iPSCs (Ren et al., 2011, Cell Res 21:849-853). The objectives of this study were to generate goat iPSCs from fetal goat primary ear fibroblasts using lentiviral transduction of four human transcription factors: OCT4, SOX2, KLF4, and cMYC. The goat iPSCs were successfully generated by co-culture with mitomycin C-treated mouse embryonic fibroblasts using medium supplemented with knockout serum replacement and human basic fibroblast growth factor. The goat iPSCs colonies are flat, compact, and closely resemble human iPSCs. They have a normal karyotype; stain positive for alkaline phosphatase, OCT4, and NANOG; express endogenous pluripotency genes (OCT4, SOX2, cMYC, and NANOG); and can spontaneously differentiate into three germ layers in vitro and in vivo.
胚胎干细胞（ESCs）是基因工程、发育生物学和疾病建模的强大模型。迄今为止，已经从老鼠（Evans 和 Kaufman，1981，《自然》292:154-156）、非人灵长类动物（Thomson 等人，Proc Nat Acad Sci USA 92:7844-7848）、人类（Thomson 等人，1998，《科学》282:1145-1147）和大鼠（Buehr 等人，Cell 135:1287-1298）中建立了 ESC；然而，从家养的有蹄类动物（如山羊、绵羊、牛和猪）中获得 ESC 的尝试尚未成功。相反，可以通过几种转录因子（OCT3/4、SOX2、KLF4、cMYC、LIN28 和 NANOG）的组合重编程体细胞来生成诱导多能干细胞（iPSCs）。迄今为止，已经从各种物种中分离出 iPSCs，但关于山羊 iPSCs 的信息有限（Ren 等人，2011，《细胞研究》21:849-853）。本研究的目的是使用四种人类转录因子（OCT4、SOX2、KLF4 和 cMYC）的慢病毒转导，从胎儿山羊原代耳成纤维细胞中生成山羊 iPSCs。山羊 iPSCs 是通过与丝裂霉素 C 处理的小鼠胚胎成纤维细胞共培养，使用含有无蛋白血清替代物和人碱性成纤维细胞生长因子的培养基成功生成的。山羊 iPSCs 集落平坦、紧凑，与人类 iPSCs 非常相似。它们具有正常的核型；碱性磷酸酶、OCT4 和 NANOG 染色阳性；表达内源性多能性基因（OCT4、SOX2、cMYC 和 NANOG）；并能在体外和体内自发分化为三个胚层。
