Wnt Inhibition Safeguards Porcine Embryonic Stem Cells From the Acquisition of Extraembryonic Endoderm Cell Fates

Abstract Porcine embryonic stem cells (ESCs) are excellent models for exploring embryogenesis, producing genetically enhanced farm animals, and improving breeding. Various chemicals have been applied to generate porcine ESCs from embryos, which differ from mouse and human ESC derivation. Wnt inhibitors XAV939 or IWR1 are required to isolate and maintain porcine ESCs. How Wnt inhibitors specify porcine ESC fate decisions remains poorly understood. Additionally, whether porcine ESCs can be converted to extraembryonic endoderm (XEN) cells without genetic interventions has not been reported. Here, it is reported that Wnt inhibitors (i.e., XAV939 and IWR1) safeguard porcine ESCs from acquiring the XEN lineage. Porcine ESCs rely on Wnt inhibitors to maintain pluripotency. Without them, porcine ESCs exit from pluripotency and convert to XEN cells. An efficient strategy and culture conditions are further developed to directly derive porcine XEN cells from ESCs without gene editing. The resulting XEN cells from ESCs exhibit similar transcriptome and chromatin accessibility features to XEN cells from embryos and contribute to mouse extraembryonic tissues. This study will deepen the understanding of porcine pluripotency, lay the foundation for deriving high‐quality porcine ESCs with germline chimerism and transmission, and provide valuable materials to study extraembryonic development and lineage segregation in livestock.