The potential of induced pluripotent stem (iPS) cells, which have self-renewal ability and can differentiate into three germ layers, led us to hypothesize that iPS cells in pigs can be useful and suitable source for producing transgenic pigs. In this study, we generated iPS-like cells using doxycycline-inducible piggyBac (PB) expression vectors encoding porcine 4 transcription factors. After transfection, transfected cells were cultured until the formation of outgrowing colonies taking least of 7-10 days. The iPS-like cells demonstrated pluripotent characteristics such as self-renewal, high proliferation, expression of pluripotent markers, and aggregation ability. The embryo development through somatic cell nuclear transfer (SCNT), cleavage rate, and blastocyst formation rate did not show any significant differences. However, the total cell number of blastocysts was significantly increased with the established cell line. In conclusion, the iPS-like cell line, generated from porcine transcriptional factors using the PB transposon system, demonstrated pluripotency with the capacity for unlimited self-renewal, and could be used as donor cells to produce cloned embryos by SCNT. These cells will be suitable for gene modification and would contribute to the stability or safety of pig models in biomedical research.

中文翻译：

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使用piggyBac系统生产由猪内源性因子诱导的多能干细胞样细胞重建的转基因猪胚。

具有自我更新能力并可以分化为三个胚层的诱导多能干（iPS）细胞的潜力使我们推测，猪中的iPS细胞可以作为生产转基因猪的有用和合适来源。在这项研究中，我们使用强力霉素诱导的piggyBac（PB）表达载体编码猪4种转录因子生成了iPS样细胞。转染后，将转染的细胞培养直至至少7-10天形成向外生长的菌落。iPS样细胞表现出多能性特征，例如自我更新，高增殖，多能性标志物的表达和聚集能力。通过体细胞核移植（SCNT），卵裂率和胚泡形成率的胚胎发育没有显示任何显着差异。然而，建立的细胞系使胚泡的总细胞数显着增加。总之，使用PB转座子系统从猪转录因子产生的iPS样细胞系表现出多能性，具有无限的自我更新能力，可以用作供体细胞，通过SCNT产生克隆的胚胎。这些细胞将适合基因修饰，并有助于生物医学研究中猪模型的稳定性或安全性。