Pig Epiblast Stem Cells Depend on Activin/Nodal Signaling for Pluripotency and Self-Renewal

Background: NANOG is a homeodomain-containing transcription factor which forms one of the hubs in the pluripotency network and plays a key role in the reprogramming of somatic cells and epiblast stem cells to naïve pluripotency. Studies have found that NANOG has many interacting partners and some of these were shown to play a role in its ability to mediate reprogramming. In this study, we set out to analyse the effect of NANOG interactors on the reprogramming process. Methods: Epiblast stem cells and somatic cells were reprogrammed to naïve pluripotency using MEK/ERK inhibitor PD0325901, GSK3β inhibitor CHIR99021 and Leukaemia Inhibitory Factor (together termed 2i Plus LIF). Zmym2 was knocked out using the CRISPR/Cas9 system or overexpressed using the PiggyBac system. Reprogramming was quantified after ZMYM2 deletion or overexpression, in diverse reprogramming systems. In addition, embryonic stem cell self renewal was quantified in differentiation assays after ZMYM2 removal or overexpression. Results: In this work, we identified ZMYM2/ZFP198, which physically associates with NANOG as a key negative regulator of NANOG-mediated reprogramming of both epiblast stem cells and somatic cells. In addition, ZMYM2 impairs the self renewal of embryonic stem cells and its overexpression promotes differentiation. Conclusions: We propose that ZMYM2 curtails NANOG’s actions during the reprogramming of both somatic cells and epiblast stem cells and impedes embryonic stem cell self renewal, promoting differentiation.

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17-P007 Activin/nodal signalling is required for self renewal of pig epiblast stem cells

Mechanisms of Development ◽

10.1016/j.mod.2009.06.728 ◽

2009 ◽

Vol 126 ◽

pp. S272-S273

Author(s):

Ramiro Alberio ◽

Nicola Croxall ◽

Cinzia Allegrucci

Keyword(s):

Stem Cells ◽

Self Renewal ◽

Epiblast Stem Cells

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The Key Role of MicroRNAs in Self-Renewal and Differentiation of Embryonic Stem Cells

International Journal of Molecular Sciences ◽

10.3390/ijms21176285 ◽

2020 ◽

Vol 21 (17) ◽

pp. 6285

Author(s):

Giuseppina Divisato ◽

Fabiana Passaro ◽

Tommaso Russo ◽

Silvia Parisi

Keyword(s):

Stem Cells ◽

Embryonic Stem Cells ◽

Cell Cycle Progression ◽

Molecular Mechanisms ◽

Developmental Stages ◽

Embryonic Stem ◽

Self Renewal ◽

Cycle Progression ◽

Epiblast Stem Cells

Naïve pluripotent embryonic stem cells (ESCs) and epiblast stem cells (EpiSCs) represent distinctive developmental stages, mimicking the pre- and the post-implantation events during the embryo development, respectively. The complex molecular mechanisms governing the transition from ESCs into EpiSCs are orchestrated by fluctuating levels of pluripotency transcription factors (Nanog, Oct4, etc.) and wide-ranging remodeling of the epigenetic landscape. Recent studies highlighted the pivotal role of microRNAs (miRNAs) in balancing the switch from self-renewal to differentiation of ESCs. Of note, evidence deriving from miRNA-based reprogramming strategies underscores the role of the non-coding RNAs in the induction and maintenance of the stemness properties. In this review, we revised recent studies concerning the functions mediated by miRNAs in ESCs, with the aim of giving a comprehensive view of the highly dynamic miRNA-mediated tuning, essential to guarantee cell cycle progression, pluripotency maintenance and the proper commitment of ESCs.

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