Wei Chao, Li Xia, Zhang Pengfei, Zhang Yu, Liu Tong, Jiang Shaoshuai, Han Fei, Zhang Yunhai
Anhui Provincial Laboratory for Local Livestock and Poultry Genetic Resource Conservation and BreedingCollege of Animal Science and Technology, Anhui Agricultural University, Hefei, Anhui 230036, China.
Anhui Provincial Laboratory for Local Livestock and Poultry Genetic Resource Conservation and BreedingCollege of Animal Science and Technology, Anhui Agricultural University, Hefei, Anhui 230036, China
Reproduction. 2015 May;149(5):485-96. doi: 10.1530/REP-14-0410. Epub 2015 Feb 2.
Partially reprogrammed induced pluripotent stem cells (PiPSCs) have great potential for investigating reprogramming mechanisms and represent an alternative potential material for making genetically modified animals and regenerative medicine. To date, PiPSCs have scarcely been reported in detail when compared with mice and humans. In this study, we obtained PiPSCs from porcine adipose-derived stem cells (pADSCs) by ectopic expression of human transcription factors (OCT4, SOX2, c-MYC, and KLF4) in feeder-free condition. The morphology and proliferation activity of porcine PiPSCs (pPiPSCs) were similar to those of porcine fully reprogrammed iPSCs (pFiPSCs); furthermore, pPiPSCs expressed higher levels of the typical surface molecules (CD29) found in pADSCs. However, pPiPSCs were negative for key proteins (NANOG) connected with stemness and possessed lower differentiation ability in vivo and in vitro. When differentiation-inhibiting factors were withdrawn, pPiPSCs-derived cells (pPiPSC-DCs) showed similar features to pADSCs in many aspects, including proliferation, differentiation, and immunosuppression. When both types of cells were used to produce cloned embryos, we found that the blastocyst formation rate of 19DC (one of the pPiPSC-DC cell lines)-derived cloned embryos was obviously higher than that of others. The total cell number of 19DC-derived blastocysts was significantly higher than the 30DC (one pFiPSC-DC cell line)-derived blastocysts. In all, through limited differentiation ability, the proliferation activity of pPiPSCs is similar to that of pFiPSCs, and pPiPSCs can retain several of the features of pADSCs, which are beneficial to cell therapy. Furthermore, the differentiation of pPiPSCs is more favorable for producing high-quality reconstructed embryos.
部分重编程诱导多能干细胞(PiPSCs)在研究重编程机制方面具有巨大潜力,并且是制造转基因动物和用于再生医学的一种潜在替代材料。迄今为止,与小鼠和人类相比,PiPSCs鲜有详细报道。在本研究中,我们通过在无饲养层条件下异位表达人类转录因子(OCT4、SOX2、c-MYC和KLF4),从猪脂肪来源干细胞(pADSCs)中获得了PiPSCs。猪PiPSCs(pPiPSCs)的形态和增殖活性与猪完全重编程的iPSCs(pFiPSCs)相似;此外,pPiPSCs表达pADSCs中发现的典型表面分子(CD29)的水平更高。然而,pPiPSCs与干性相关的关键蛋白(NANOG)呈阴性,并且在体内和体外具有较低的分化能力。当去除分化抑制因子时,pPiPSCs衍生细胞(pPiPSC-DCs)在许多方面表现出与pADSCs相似的特征,包括增殖、分化和免疫抑制。当使用这两种类型的细胞来生产克隆胚胎时,我们发现19DC(一种pPiPSC-DC细胞系)衍生的克隆胚胎的囊胚形成率明显高于其他胚胎。19DC衍生囊胚的总细胞数显著高于30DC(一种pFiPSC-DC细胞系)衍生的囊胚。总之,通过有限的分化能力,pPiPSCs的增殖活性与pFiPSCs相似,并能保留pADSCs的一些特征,这有利于细胞治疗。此外,pPiPSCs的分化更有利于产生高质量的重构胚胎。