State Key Laboratory for Agrobiotechnology, College of Biological Sciences, China Agricultural University, Beijing, China.
Sci China Life Sci. 2011 Jun;54(6):553-9. doi: 10.1007/s11427-011-4179-5. Epub 2011 Jun 26.
Induced pluripotent stem (iPS) cell technology demonstrates that somatic cells can be reprogrammed to a pluripotent state by over-expressing four reprogramming factors. This technology has created an interest in deriving iPS cells from domesticated animals such as pigs, sheep and cattle. Moloney murine leukemia retrovirus vectors have been widely used to generate and study mouse iPS cells. However, this retrovirus system infects only mouse and rat cells, which limits its use in establishing iPS cells from other mammals. In our study, we demonstrate a novel retrovirus strategy to efficiently generate porcine iPS cells from embryonic fibroblasts. We transfected four human reprogramming factors (Oct4, Sox2, Klf4 and Myc) into fibroblasts in one step by using a VSV-G envelope-coated pantropic retrovirus that was easily packaged by GP2-293 cells. We established six embryonic stem (ES)-like cell lines in human ES cell medium supplemented with bFGF. Colonies showed a similar morphology to human ES cells with a high nuclei-cytoplasm ratio and phase-bright flat colonies. Porcine iPS cells could form embryoid bodies in vitro and differentiate into the three germ layers in vivo by forming teratomas in immunodeficient mice.
诱导多能干细胞（iPS）技术表明，通过过表达四种重编程因子，体细胞可以被重新编程为多能状态。这项技术激发了人们从猪、绵羊和牛等家畜中提取 iPS 细胞的兴趣。莫洛尼鼠白血病逆转录病毒载体已被广泛用于生成和研究小鼠 iPS 细胞。然而，这种逆转录病毒系统仅感染小鼠和大鼠细胞，这限制了它在建立其他哺乳动物的 iPS 细胞中的应用。在我们的研究中，我们展示了一种从胚胎成纤维细胞中高效生成猪 iPS 细胞的新型逆转录病毒策略。我们通过使用一种由 GP2-293 细胞容易包装的、带有 VSV-G 包膜的泛嗜性逆转录病毒，一步转染四个人类重编程因子（Oct4、Sox2、Klf4 和 Myc）到成纤维细胞中。我们在含有 bFGF 的人胚胎干细胞培养基中建立了六个类胚胎干细胞（ES）细胞系。集落表现出与人类 ES 细胞相似的形态，具有高核质比和明亮的扁平集落。猪 iPS 细胞可以在体外形成胚状体，并通过在免疫缺陷小鼠中形成畸胎瘤在体内分化为三个胚层。
