Zhao Lixia, Wang Zixin, Zhang Jindun, Yang Jian, Gao Xuefei, Wu Baojiang, Zhao Gaoping, Bao Siqin, Hu Shuxiang, Liu Pentao, Li Xihe
Research Center For Animal Genetic Resources of Mongolia Plateau, Inner Mongolia University, Hohhot, 010021, China; Inner Mongolia Saikexing Institute of Breeding and Reproductive Biotechnology in Domestic Animal, Hohhot, 011517, China.
Inner Mongolia Saikexing Institute of Breeding and Reproductive Biotechnology in Domestic Animal, Hohhot, 011517, China.
Tissue Cell. 2017 Oct;49(5):521-527. doi: 10.1016/j.tice.2017.05.005. Epub 2017 May 22.
Single-cell derived bovine induced pluripotent stem cells (iPSCs) were generated by the introduction of piggyBac transposons with CAG promoting transcription factors (Oct3/4, Sox2, Klf4 and cMyc). In the study, the bovine iPSCs colony from single cell could passage more than 50 passages after enzymatic dissociation into single cells. These bovine iPSCs cells kept the normal karyotype and displayed dome shaped clones similar to mouse embryonic stem cells. They showed pluripotency in many ways, including their expression of pluripotency markers, such as OCT3/4, NANOG, SOX2, SSEA1, SSEA4, and AP in immunofluorescence assay, Oct4, Nanog, Sox2, Klf4 and cMyc in RT-PCR. Additionally, single-cell derived bovine iPSCs formed embryoid bodies and teratomas that all subsequently gave rise to differentiated cells from all three embryonic germ layers. The results showed that our reprogramming method could obtain high efficiency single-cell cloning bovine iPSCs, and the efficiency of single cell cloning is 40%.
通过导入携带CAG启动转录因子（Oct3/4、Sox2、Klf4和cMyc）的猪尾巴转座子，产生了单细胞来源的牛诱导多能干细胞（iPSCs）。在该研究中，单细胞来源的牛iPSCs集落经酶解为单细胞后可传代50代以上。这些牛iPSCs细胞保持正常核型，并呈现出与小鼠胚胎干细胞相似的穹顶状克隆。它们在许多方面表现出多能性，包括在免疫荧光分析中表达多能性标志物，如OCT3/4、NANOG、SOX2、SSEA1、SSEA4和AP，在RT-PCR中表达Oct4、Nanog、Sox2、Klf4和cMyc。此外，单细胞来源的牛iPSCs形成了胚状体和畸胎瘤，随后均分化出所有三个胚胎胚层的细胞。结果表明，我们的重编程方法能够高效获得单细胞克隆牛iPSCs，单细胞克隆效率为40%。
