|Junjun Xu,Hongjiang Wei and Jianyong Han et al.Generation of pig induced pluripotent stem cells using an extended pluripotent stem cell culture system. Stem Cell Research & Therapy doi.org/10.1186/s13287-019-1303-0|
.Generation of pig induced pluripotent stem cells using an extended pluripotent stem cell culture system.
Junjun Xu,Leqian Yu,Jianxiong Guo,Jinzhu Xiang,Zheng Zheng,Dengfeng Gao,Bingbo Shi,Haiyang Hao,Deling Jiao,Liang Zhong,Yu Wang,Jun Wu,Hongjiang Wei and Jianyong Han
Stem Cell Research & Therapy
Background: Pigs have emerged as one of the most popular large animal models in biomedical research, which in many cases is considered as a superior choice over rodent models. In addition, transplantation studies using pig pluripotent stem (PS) cell derivatives may serve as a testbed for safety and efficacy prior to human trials. Recently, it has been shown that mouse and human PS cells cultured in LCDM (recombinant human LIF, CHIR 99021, (S)-(+)-dimethindene maleate, minocycline hydrochloride) medium exhibited extended developmental potential (designated as extended pluripotent stem cells, or EPS cells), which could generate both embryonic and extraembryonic tissues in chimeric mouse conceptus. Whether stable pig induced pluripotent stem (iPS) cells can be generated in LCDM medium and their chimeric competency remains unknown. Methods: iPS cells were generated by infecting pig pericytes (PC) and embryonic fibroblasts (PEFs) with a retroviral vector encoding Oct4, Sox2, Klf4, and cMyc reprogramming factors and subsequently cultured in a modified LCDM medium. The pluripotency of PC-iPS and PEF-iPS cells was characterized by examining the expression of pluripotencyrelated transcription factors and surface markers, transcriptome analysis, and in vitro and in vivo differentiation capabilities. Chimeric contribution of PC-iPS cells to mouse and pig conceptus was also evaluated with fluorescence microscopy, flow cytometry, and PCR analysis.