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EMT blockage is required for mouse naïve pluripotent stem cell derivation

ABSTRACT: Pluripotency is the differentiation capacity of particular cells exhibited in the early embryo in vivo and embryonic stem (ES) cells have been shown to originate from the inner cell mass (ICM) of an E3.5 blastocyst. Although the potential for ES cells to differentiate into the three germ layers is equated to ICM cells, they differ in the ability to maintain the capacity for self-renewal. Despite several studies on the maintenance of ES cells in the ground state of pluripotency, the precise mechanism of conversion from the ICM to the ES cell remains unclear. Here , we have examined the cell characteristics and expression profile within the intermediate stages of ES cell derivation from the ICM. Gene clustering and ontology (GO) analyses showed a significant change in the expression of epigenetic modifiers and DNA methylation-related genes in the intermediate stages. We have proposed that an epithelial-to-mesenchymal transition (EMT) blockage is required during derivation of mouse ES cells from E3.5 blastocysts. This study suggests a novel mechanistic insight into ES cell derivation and provides a time-course transcriptome profiling resource for the dissection of gene regulatory networks that underlie the transition from ICM to ES cells.

ORGANISM(S): Mus musculus

PROVIDER: GSE87793 | GEO | 2017/10/02

SECONDARY ACCESSION(S): PRJNA347627

REPOSITORIES: GEO

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Multifaceted SOX2-chromatin interaction underpins pluripotency progression in early embryos

Project description:Pioneer transcription factors (TFs), such as OCT4 and SOX2, play crucial roles in pluripotency regulation. However, the master TF-governed pluripotency regulatory circuitry was largely inferred from cultured cells. In this work, we investigated SOX2 binding from embryonic day 3.5 (E3.5) to E7.5 in the mouse. In E3.5 inner cell mass (ICM), SOX2 regulates the ICM-trophectoderm program but is dispensable for opening global enhancers. Instead, SOX2 occupies preaccessible enhancers in part opened by early-stage expressing TFs TFAP2C and NR5A2. SOX2 then redistributes when cells adopt naive and formative pluripotency by opening enhancers or poising them for further rapid activation. Hence, multifaceted pioneer TF–enhancer interaction underpins pluripotency progression in embryos, including a state in E3.5 ICM that bridges totipotency and pluripotency.

2023-12-15 | GSE203194 | GEO

Transcription profiling of mouse inner cell mass (ICM) from a single cells derived from mouse blastocysts at E3.5

Project description:The inner cell mass (ICM) of the early blastocyst at E3.5, a source of ES cell derivation, is a morphologically homogeneous population of undifferentiated pluripotent cells that give rise to all embryonic lineages. The immediate application of the newly developed V1V3 method to single cells in this stage of mouse embryos revealed the presence of two populations of cells, one with primitive endoderm expression and the other with pluripotent epiblast-like gene expression. The genes expressed differentially between these two populations were well preserved in morphologically differentiated primitive endoderm and epiblast in the embryos one day later (E4.5), demonstrating that the method successfully detects subtle but essential differences in gene expression at the single-cell level among seemingly homogeneous cell populations. This study provides a strategy to analyze biophysical events in medicine as well as in neural, stem cell, and developmental biology, where small numbers of distinctive or diseased cells play critical roles. Experiment Overall Design: We isolated blastocysts at E3.5 and dissociated the ICM into single cells by trypsin-EDTA treatment. To prepare cDNA samples, we then randomly picked a total of 55 single cells. cDNAs were synthesized and amplified by the V1V3 method, and screened by gene-specific PCR using Oct4 and Cdx2 to remove trophectoderm cells, and 50 cells were identified as Oct4-positive and Cdx2-negative, ICM cells.

2008-06-13 | E-GEOD-4307 | biostudies-arrayexpress

Transcriptional landscape changes during human embryonic stem cell derivation

Project description:Currently, there is only minimal information elucidating the transcriptional changes occurring during the in vitro transition of the inner cell mass (ICM) towards the human embryonic stem cell (hESC) stage and the role played by the post inner cell mass intermediate (PICMI). In this study, we perform in-depth analysis of the transcriptional heterogeneity between these three stages of hESC derivation in order to correlate their downstream effects on pluripotency states and differentiation. We reveal that although PICMI is transcriptionally in close proximity to the hESC profile and distinct from ICM, it exhibits upregulation of primordial germ cell markers, dependence on LIF signaling, upregulation of naïve pluripotency-specific signaling networks and appears to be a switching point from naïve to primed pluripotency. This indicates similarities with mouse epiblast-like cells displaying a transient state of pluripotency, between naïve and primed pluripotent states. Overall, we highlight the need to gain insight into the molecular and transcriptional landscape during hESC derivation, which will further help in the derivation of human PGCs and naïve hESCs in vitro which remains inefficient.

2018-09-24 | GSE119378 | GEO

Metastable pluripotent states in NOD mouse derived ES cells

Project description:Embryonic stem (ES) cells are isolated from the inner cell mass (ICM) of developing blastocysts, whereas epiblast stem cells (EpiSCs) are derived from the post-implantation epiblast and are characterized by a restricted developmental potential. Although certain mouse strains, such as the non-obese diabetic (NOD) mice, are considered “non-permissive” for ES cell derivation, they retain the capacity to generate EpiSCs. Using the NOD strain as a model, we characterized the stability of pluripotent states in cells generated by ICM explantation or direct in vitro reprogramming. We find that ES-like NOD stem cells can be captured in both approaches by providing exogenous constitutive expression of Klf4 or c-Myc transcription factors or small molecules that can replace these factors during in vitro reprogramming. The fully pluripotent NOD ES and iPS cells appear “metastable”, as the cells acquire an alternative EpiSC-like identity after removal of the exogenous factors, while reintroduction of these factors converts the cells back to ICM-like pluripotency. Our findings suggest that stem cells from different genetic backgrounds can assume distinct states of pluripotency in vitro, the stability of which is regulated by endogenous genetic determinants and can be modified by the continuous presence of defined exogenous factors. Gene expression profiling was performed in mouse ES, EpiSC and EpiSC-like cell lines.

2009-04-09 | E-GEOD-15603 | biostudies-arrayexpress

Establishment of trophoblast stem cells under defined culture conditions in mice

Project description:The inner cell mass (ICM) and trophoblast cell lineages duet early embryonic development in mammals. After implantation, the ICM forms the embryo proper as well as some extraembryonic tissues, whereas the trophoectoderm (TE) exclusively forms the fetal portion of the placenta and the trophoblast giant cells. Although embryonic stem (ES) cells can be derived from ICM in cultures of mouse blastocysts in the presence of LIF and/or combinations of small-molecule chemical compounds, and the undifferentiated pluripotent state can be stably maintained without use of serum and feeder cells, defined culture conditions for derivation and maintenance of undifferentiated trophoblast stem (TS) cells have not been established. Here, we report that addition of FGF2, activin A, XAV939, and Y27632 are necessary and sufficient for derivation of TS cells from both of E3.5 blastocysts and E6.5 early postimplantation extraembryonic ectoderm. Moreover, the undifferentiated TS cell state can be stably maintained in chemically defined culture conditions. Cells derived in this manner expressed TS cell marker genes, including Eomes, Elf5, Cdx2, Klf5, Cdh1, Esrrb, Sox2, and Tcfap2c; differentiated into all trophoblast subtypes (trophoblast giant cells, spongiotrophoblast, and labyrinthine trophoblast) in vitro; and exclusively contributed to trophoblast lineages in chimeric animals. This delineation of minimal requirements for derivation and self-renewal provides a defined platform for precise description and dissection of the molecular state of TS cells.

2014-07-08 | GSE59107 | GEO

Establishment of trophoblast stem cells under defined culture conditions in mice

2014-07-08 | E-GEOD-59107 | biostudies-arrayexpress

Metastable pluripotent states in NOD mouse derived ES cells

2009-04-09 | GSE15603 | GEO

Genome-wide Promoter Mapping of Histone H3 Lysine 4 and Lysine 27 Methylation in Preimplantation Embryos

Project description:First lineage specification in the mammalian blastocyst embryo leads to formation of the inner cell mass (ICM) and trophectoderm (TE), which respectively give rise to the embryo proper and extraembryonic tissues. We show histone methylation asymmetry on promoters in the first two developmental lineages, and highlight epigenetic skewing associated with derivation of embryonic stem (ES) cells. Comparison of histone methylation patterns on promoters in the ICM, TE and in ES cells derived from ICM. ChIP-chip experiments using anti-H3K4me3 or anti-H3K27me3 antibodies. Two or three replicates per sample.

2010-05-14 | E-GEOD-17387 | biostudies-arrayexpress

Expression data from single cells from ICMs of mouse blastocysts at E3.5

2006-04-26 | GSE4307 | GEO

RNA sequencing of mouse embryonic day 3.5 blastocyst inner cell mass cells

Project description:Inner cell mass (ICM) cells of two independent E3.5 blastocysts were profiled by transcriptome sequencing to assess the expression of extracellular matrix components.

2014-05-15 | E-MTAB-2555 | biostudies-arrayexpress

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