Characterization of porcine induced pluripotent stem cells derived from Sertoli cells with activation of Wnt signaling pathway. Characterization of porcine induced pluripotent stem cells derived from Sertoli cells with activation of Wnt signaling pathway

Studies on porcine induced pluripotent stem (iPS) cells served as a great reference for human clinical research due to the similarity in organ size, physiology, anatomy, and heredity between pig and human. However, the reprogramming efficiency of iPS cells was currently poor. In this study, we reported the induction of iPS cells from porcine Sertoli cells (SCs) via infecting with four separate lentiviral vectors expressing pig OCT4, SOX2, KLF4 and c-MYC. Typical embryonic stem-like cells began to form on day 3 after infection, and alkaline phosphatase (AP)-positive colonies were picked up on 7 days. The porcine iPS cells derived from SCs, termed SC-iPS cells, strongly expressed pluripotent markers, showed a normal karyotype, and had differentiation characteristics to the three germ layers and primordial germ cell-like cells via an incipient mesoderm-like cell state. Further results showed that OCT4 was the core factor for AP-positive colonies formation, and the OCT4 and c-MYC were the minimum combination group to achieve the SCs reprogramming process. RNA-seq results showed that SCs could express BMP4 and activate the Wnt signaling pathway, which promoted reprogramming process. In conclusion, our findings demonstrated that the SCs could efficiently serve as a better cell source for reprogramming than porcine embryonic fibroblasts and lay a foundation of achieving the trans-differentiation capacity from SCs to germ cells. Overall design: RNA-seq in SCs, SCs-piPSCs (iPS cells derived from Sertoli Cells), PEF-piPSCs (iPS cells derived from embryonic fibroblasts).