Epiblast Stem Cell Subpopulations Represent Mouse Embryos of Distinct Pregastrulation Stages

Han, Dong Wook; Tàpia, Natalia; Joo, Jin Young; Greber, Boris; Araúzo-Bravo, Marcos J.; Bernemann, Christof; Ko, Kinarm; Wu, Guangming; Stehling, Martin; Tae, Jeong; Schöler, Hans R.

doi:10.1016/j.cell.2010.10.015

Cited by 192 publications

(214 citation statements)

References 27 publications

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“…In the post‐implantation epiblast, NANOG is re‐expressed (Hart et al , 2004; Osorno et al , 2012; Hoffman et al , 2013) but ESRRB is not (Adachi et al , 2013), likely due to differences in signaling environments between pre‐ and post‐implantation epiblasts. It is noteworthy that subpopulations of ESC cultures have been proposed to bear a similar character to primed pluripotent cells and vice versa (Hayashi et al , 2008; Han et al , 2010). Upon differentiation, naïve ESCs transit through a state (named EpiLCs) with transcriptional similarity to the early post‐implantation epiblast (Buecker et al , 2010; Hayashi et al , 2011).…”

Section: Discussionmentioning

confidence: 99%

Esrrb extinction triggers dismantling of naïve pluripotency and marks commitment to differentiation

et al. 2018

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Self‐renewal of embryonic stem cells (ESCs) cultured in LIF/fetal calf serum (FCS) is incomplete with some cells initiating differentiation. While this is reflected in heterogeneous expression of naive pluripotency transcription factors (TFs), the link between TF heterogeneity and differentiation is not fully understood. Here, we purify ESCs with distinct TF expression levels from LIF/FCS cultures to uncover early events during commitment from naïve pluripotency. ESCs carrying fluorescent Nanog and Esrrb reporters show Esrrb downregulation only in Nanoglow cells. Independent Esrrb reporter lines demonstrate that Esrrbnegative ESCs cannot effectively self‐renew. Upon Esrrb loss, pre‐implantation pluripotency gene expression collapses. ChIP‐Seq identifies different regulatory element classes that bind both OCT4 and NANOG in Esrrbpositive cells. Class I elements lose NANOG and OCT4 binding in Esrrbnegative ESCs and associate with genes expressed preferentially in naïve ESCs. In contrast, Class II elements retain OCT4 but not NANOG binding in ESRRB‐negative cells and associate with more broadly expressed genes. Therefore, mechanistic differences in TF function act cumulatively to restrict potency during exit from naïve pluripotency.

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Section: Discussionmentioning

confidence: 99%

Esrrb extinction triggers dismantling of naïve pluripotency and marks commitment to differentiation

et al. 2018

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“…The bisulfite‐converted DNA was amplified by PCR with previously described primers 70, 71. The PCR products were cloned into the pCRII TOPO vector (Invitrogen), and plasmids extracted from individual clones were sequenced by GATC‐biotech (http://www.gatc-biotech.com/en/index.html).…”

Section: Methodsmentioning

confidence: 99%

“…Eight‐cell embryos were flushed from female mice at E2.5 and placed in M2 medium 70. Trypsin‐digested iPSCs (8–12 cells and one embryo per aggregate) were transferred into a depression in the microdrop of KSOM.…”

Section: Methodsmentioning

confidence: 99%

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Changing POU dimerization preferences converts Oct6 into a pluripotency inducer

Jerabek

et al. 2016

EMBO Reports

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123

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The transcription factor Oct4 is a core component of molecular cocktails inducing pluripotent stem cells (iPSCs), while other members of the POU family cannot replace Oct4 with comparable efficiency. Rather, group III POU factors such as Oct6 induce neural lineages. Here, we sought to identify molecular features determining the differential DNA‐binding and reprogramming activity of Oct4 and Oct6. In enhancers of pluripotency genes, Oct4 cooperates with Sox2 on heterodimeric SoxOct elements. By re‐analyzing ChIP‐Seq data and performing dimerization assays, we found that Oct6 homodimerizes on palindromic OctOct more cooperatively and more stably than Oct4. Using structural and biochemical analyses, we identified a single amino acid directing binding to the respective DNA elements. A change in this amino acid decreases the ability of Oct4 to generate iPSCs, while the reverse mutation in Oct6 does not augment its reprogramming activity. Yet, with two additional amino acid exchanges, Oct6 acquires the ability to generate iPSCs and maintain pluripotency. Together, we demonstrate that cell type‐specific POU factor function is determined by select residues that affect DNA‐dependent dimerization.

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“…Luciferase constructs containing the proximal enhancer (PE) and the distal enhancer (DE) of the Oct4 promoter [4,13] were transfected into EpiSCs and mESCs, along with renilla normalization constructs, using Lipofectamine 2000 (Invitrogen). Cells were harvested 48 hours after transfection and processed according to the DualGlo Luciferase protocol (Promega, Madison, Wisconsin, www.promega.com).…”

Section: Luciferase Assaysmentioning

confidence: 99%

Distinct Developmental Ground States of Epiblast Stem Cell Lines Determine Different Pluripotency Features

et al. 2011

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Epiblast stem cells (EpiSCs) are pluripotent stem cells derived from mouse postimplantation embryos at embryonic day (E) 5.5-E7.5 at the onset of gastrulation, which makes them a valuable tool for studying mammalian postimplantation development in vitro. EpiSCs can also be reprogrammed into a mouse embryonic stem cell (mESC)-like state. Some reports have shown that the reversion of EpiSCs requires transcription factor overexpression, whereas others have suggested that use of stringent mESC culture conditions alone is sufficient for the reversion of EpiSCs. To clarify these discrepancies, we systematically compared a panel of independent EpiSC lines. We found that-regardless of the embryonic day of derivation-the different EpiSC lines shared a number of defining characteristics such as the ability to form teratomas. However, despite use of standard EpiSC culture conditions, some lines exhibited elevated expression of genes associated with mesendodermal differentiation. Pluripotency (Oct4) and mesodermal (Brachyury) marker genes were coexpressed in this subset of lines. Interestingly, the expression of mesendodermal marker genes was negatively correlated with the cells' ability to efficiently undergo neural induction. Moreover, these mesodermal marker gene-expressing cell lines could not be efficiently reverted to an mESC-like state by using stringent mESC culture conditions. Conversely, Brachyury overexpression diminished the reversion efficiency in otherwise Brachyury-negative lines. Overall, our data suggest that different EpiSC lines may undergo self-renewal into distinct developmental states, a finding with important implications for functional readouts such as reversion of EpiSCs to an mESC-like state as well as directed differentiation. STEM CELLS

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Epiblast Stem Cell Subpopulations Represent Mouse Embryos of Distinct Pregastrulation Stages

Cited by 192 publications

References 27 publications

Esrrb extinction triggers dismantling of naïve pluripotency and marks commitment to differentiation

Esrrb extinction triggers dismantling of naïve pluripotency and marks commitment to differentiation

Changing POU dimerization preferences converts Oct6 into a pluripotency inducer

Distinct Developmental Ground States of Epiblast Stem Cell Lines Determine Different Pluripotency Features

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