Pluripotent stem cells (PSCs) are in vitro adaptations of in vivo pluripotency continuum and can be broadly classified into naïve state characteristic of pre-implantation epiblast and primed state resembling peri-gastrulation epiblasts. Naïve and primed PSCs differ in their cellular and molecular characteristics, e.g., molecular mechanisms for maintaining undifferentiated state. Naïve-to-primed PSC transition provides a tractable in vitro model to study pluripotency development in vivo. We previously developed a protocol that enabled high-efficient (100%) and homogenous derivation of ground state of primed epiblast stem cells (rsEpiSCs) by culturing the isolated post-implantation mouse epiblast under the culture condition containing FGF2 and a Wnt signaling inhibitor (IWR1) (F/R1 condition). Based on F/R1 condition, in this study, we developed three naïve-to-primed conversion methods for generating rsEpiSCs from naïve ground state of mouse ESCs (2i/LIF condition). We found that stepwise methods, but not directly, were effective for bona fide rsEpiSCs conversion from mouse ESCs. In sum, we established a robust and efficient ground states of naïve-to-primed PSC conversion strategy that will facilitate the study of genetic, epigenetic and metabolic processes involved in pluripotency progression in vivo.
Keywords: Conversion; Ground state; Naïve; Primed; p53; rsEpiSC.
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