Reprogramming of Sheep Fibroblasts into Pluripotency under a Drug-Inducible Expression of Mouse-Derived Defined Factors
Reprogramming of Sheep Fibroblasts into Pluripotency under a Drug-Inducible Expression of Mouse-Derived Defined Factors
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DOI：
10.1371/journal.pone.0015947
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发表时间：
2011-01-06
期刊：
影响因子：
3.7
通讯作者：
Liu, Dongjun
中科院分区：
文献类型：
--
作者：
Li, Yang;Cang, Ming;Liu, Dongjun
动物胚胎干细胞（ESCs）为早期胚胎发育、基因靶向、克隆和再生医学的研究提供了强有力的工具。然而，大多数从大型动物，特别是有蹄类哺乳动物身上建立ESC系的尝试都失败了。最近，另一种类型的多能干细胞，即诱导多能干细胞（iPSCs），已经成功地从小鼠、人、猴、大鼠和猪身上产生。在这项研究中，我们展示了绵羊成纤维细胞可以通过使用药物诱导系统的定义因子重新编程为多能性。以这种方式获得的绵羊iPSCs具有正常的核型，表现出与人类ESCs相似的形态学特征，并表达AP、Oct4、Sox2、Nanog和细胞表面标记物SSEA-4。胚胎样体（EB）和畸胎瘤形成实验进一步证实了这些细胞的多能性，并产生了所有三种胚层的衍生物。我们的研究结果还表明，在培养中用胎牛血清替代敲除血清（KSR）可以提高绵羊iPSCs的重编程效率。绵羊iPSCs的产生使绵羊处于大型动物临床前试验和涉及动物基因组修改的实验的前沿。
Animal embryonic stem cells (ESCs) provide powerful tool for studies of early embryonic development, gene targeting, cloning, and regenerative medicine. However, the majority of attempts to establish ESC lines from large animals, especially ungulate mammals have failed. Recently, another type of pluripotent stem cells, known as induced pluripotent stem cells (iPSCs), have been successfully generated from mouse, human, monkey, rat and pig. In this study we show sheep fibroblasts can be reprogrammed to pluripotency by defined factors using a drug-inducible system. Sheep iPSCs derived in this fashion have a normal karyotype, exhibit morphological features similar to those of human ESCs and express AP, Oct4, Sox2, Nanog and the cell surface marker SSEA-4. Pluripotency of these cells was further confirmed by embryoid body (EB) and teratoma formation assays which generated derivatives of all three germ layers. Our results also show that the substitution of knockout serum replacement (KSR) with fetal bovine serum in culture improves the reprogramming efficiency of sheep iPSCs. Generation of sheep iPSCs places sheep on the front lines of large animal preclinical trials and experiments involving modification of animal genomes.