Associate Professor Huazhong Agricultural University Wuhan, Hubei, China
Abstract: The molecular characteristics of expanded pluripotent stem cells (EPSCs) have been a subject of debate. Human EPSCs (hEPSCs) are akin to eight-cell and morula-stage embryos, whereas mouse EPSCs (mEPSCs) resemble post-implantation epiblasts. This study investigates the developmental potential and molecular trait of porcine EPSCs (pEPSCs), revealing their unique transcriptional heterogeneity and similarities to both morula and post-implantation epiblast stages. Through single-cell transcriptomic analysis, we identified three distinct subpopulations (C1, C2, and C3) corresponding to totipotency, naïve pluripotency, and primed pluripotency. The use of dual fluorescence reporter system demonstrated transitions between these states, mirroring embryonic development. Mechanistically, we established a framework to understand the conserved roles of OTX2 and LEUTX in regulating transitions between totipotency and pluripotency, both in cells and early embryos. OTX2 promotes the transition to pluripotency by activating pluripotency and lineage priming genes, while LEUTX enhances totipotency and supports blastoid formation. This study bridges gaps in our understanding of mammalian stem cells and underscores the potential of pEPSCs in agricultural and biomedical applications.
