Epiblast stem cells (EpiSCs) are pluripotent stem cells derived from epiblasts of postimplantation mouse embryos, and thus provide a usefulmodel for studying ‘‘primed’’ pluripotent states. Here, we devised a simple and robust technique to derive high-quality EpiSCs using aninhibitor of WNT secretion. Using this method, we readily established EpiSC lines with high efficiency and were able to use whole embryonicportions without having to separate the epiblast from the visceral endoderm (VE). Expression analyses revealed that these EpiSCsmaintained a homogeneous, undifferentiated status, yet showed high potential for differentiation both in vitro and in teratomas. UnlikeEpiSCs derived by the original protocol, new EpiSC lines required continuous treatment with the Wnt inhibitor, suggesting someintrinsic differences from the existing EpiSCs. The homogeneous properties of this new version of EpiSCs should facilitate studies onthe establishment and maintenance of a ‘‘primed’’ pluripotent state, and directed differentiation from the primed state.
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