A simple and robust method for establishing homogeneous mouse epiblast stem cell lines by wnt inhibition.
Bottom Line: Here, we devised a simple and robust technique to derive high-quality EpiSCs using an inhibitor of WNT secretion.Expression analyses revealed that these EpiSCs maintained a homogeneous, undifferentiated status, yet showed high potential for differentiation both in vitro and in teratomas.The homogeneous properties of this new version of EpiSCs should facilitate studies on the establishment and maintenance of a "primed" pluripotent state, and directed differentiation from the primed state.
Affiliation: Technology and Development Team for Mammalian Genome Dynamics, RIKEN BioResource Center, 3-1-1 Koyadai, Tsukuba, Ibaraki 305-0074, Japan.Show MeSH
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Mentions: When IWP-2 was withdrawn from the EpiSC culture medium, our EpiSCs changed their morphologies and started to differentiate into cells expressing the mesoderm marker T or the endoderm marker GATA4 (Figures 3A, 3B, and S2). By contrast, 129C1, the EpiSC line established by the original protocol, could be maintained as an undifferentiated form in the absence of IWP-2. Flow-cytometry analysis of SSEA1-positive cells indicated that our EpiSC line downregulated SSEA1 expression, whereas a higher percentage of 129C1 cells continued to express SSEA1 4 days after IWP-2 removal (Figure S2).
Affiliation: Technology and Development Team for Mammalian Genome Dynamics, RIKEN BioResource Center, 3-1-1 Koyadai, Tsukuba, Ibaraki 305-0074, Japan.