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Effect of L-Proline on mouse embryonic stem cells (ESCs)

ABSTRACT: We found that the non-essential amino acid L-Proline (L-Pro) acts as a signaling molecule that promotes the conversion of embryonic stem cells (ESCs) into mesenchymal-like, spindle-shaped, highly motile, invasive pluripotent stem cells. This embryonic stem cell-to-mesenchymal-like transition (esMT) is accompanied by a genome-wide remodeling of the transcriptome We used microarrays to elucidate whether a diverse transcriptional program is the basis of the morphological and motility differences between ESCs and L- Proline treated ESCs (PiCs) Total RNA was extracted from Control (ESCs) and L-Proline treated mouse embryonic stem cells (PiCs) and hybridized on Affimetrix microarrays.

ORGANISM(S): Mus musculus

SUBMITTER: Gabriella Minchiotti

PROVIDER: E-GEOD-50603 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Effect of L-Proline on mouse embryonic stem cells (ESCs)

Project description:We found that the non-essential amino acid L-proline (L-Pro) acts as a signaling molecule that promotes the conversion of embryonic stem cells (ESCs) into mesenchymal-like, spindle-shaped, highly motile, invasive pluripotent stem cells.This embryonic stem cell-to-mesenchymal-like transition (esMT) is accompanied by a genome-wide remodeling of the H3K9me3 and H3K36me3 histone marks. Examination of 2 different histone modifications in untreated ESCs and L-Pro treated ESCs

2013-08-31 | E-GEOD-50477 | biostudies-arrayexpress

Effect of L-Proline on mouse embryonic stem cells (ESCs)

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2013-09-05 | GSE50603 | GEO

REX1 is the critical target of RNF12 in imprinted X chromosome inactivation in mice.

Project description:In mice, imprinted X-chromosome inactivation (iXCI) of the paternal X in the pre-implantation embryo and extra-embryonic tissues is followed by X-reactivation in the epiblast precursors of the inner cell mass (ICM) of the blastocyst, to facilitate initiation of random XCI (rXCI) in all embryonic tissues. RNF12 is an E3 ubiquitin-ligase that plays a key role in XCI. RNF12 targets pluripotency protein REX1 for degradation to initiate rXCI in embryonic stem cells (ESCs) and loss of the maternal copy of Rnf12 leads to embryonic lethality due to iXCI failure. Here, we show that loss of Rex1 rescues the rXCI phenotype observed in Rnf12-/- ESCs, and that REX1 is the prime target of RNF12 in ESCs. Genetic ablation of Rex1 in Rnf12-/- mice rescues the Rnf12-/- iXCI phenotype, and results in viable and fertile Rnf12-/-:Rex1-/- female mice displaying normal iXCI and rXCI. Our results show that REX1 is the critical target of RNF12 in XCI.

2018-11-15 | PXD010944 | Pride

Effect of L-Proline on mouse embryonic stem cells (ESCs)

2013-08-31 | GSE50477 | GEO

Quantitative subcellular proteomics using SILAC reveals enhanced metabolic buffering in ground state pluripotency

Project description:The ground state of pluripotency is defined as a minimal unrestricted state as present in the Inner Cell Mass (ICM). Mouse embryonic stem cells (ESCs) grown in a defined serum-free medium with two kinase inhibitors (‘2i’) reflect this state, whereas ESCs grown in the presence of serum (‘serum’) share more similarities with post implantation epiblast cells. Pluripotency results from an intricate interplay between cytoplasmic, nuclear and chromatin-associated proteins. Therefore, quantitative information on the (sub)cellular proteome is essential to gain insight in the molecular mechanisms driving different pluripotent states. Here, we describe a full SILAC workflow and quality controls for proteomic comparison of 2i and serum ESCs. We demonstrate that this workflow is applicable for subcellular proteomics of the cytoplasm, nuclear and chromatin. The obtained quantitative information revealed increased levels of naïve pluripotency factors on the chromatin of 2i ESCs. Further, we demonstrate that these pluripotent states are supported by distinct metabolic programs, which include upregulation of free radical buffering by the glutathione pathway in 2i ESCs. Through induction of intracellular radicals, we show that the altered metabolic environment renders 2i ESCs less sensitive to oxidative stress. Altogether, this work provides novel insights into the proteome landscape underlying ground state pluripotency.

2018-11-07 | PXD009217 | Pride

Ssrp1 co-immunoprecipitation and mass spectrometry

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2020-08-24 | PXD017351 | Pride

A mass spectrometry survey of chromatin-associated proteins in pluripotency and early lineage commitment

Project description:Pluripotent Embryonic Stem Cells (ESCs) can be captured in vitro in different states, ranging from unrestricted ‘naïve’ to more developmentally constrained ‘primed’ pluripotency. Complexes involved in epigenetic regulation and key transcription factors have been shown to be involved in specifying these distinct states. In this study, we use proteomic profiling of the chromatin landscape in naive pluripotent ESCs, Epistem cells (EpiSCs) and early differentiated ESCs to survey the chromatin in naïve and primed pluripotency and during differentiation. We provide a comprehensive overview of epigenetic complexes situated on the chromatin and identify proteins associated with the maintenance and loss of pluripotency. The findings presented here indicate major compositional alterations of epigenetic complexes starting from ESC priming onwards. Our results contribute to the understanding of ESC differentiation and provide a framework for future studies of lineage commitment of ESCs.

2019-07-04 | PXD011782 | Pride

Investigating the role of Lef1 in mouse embryonic stem cells.

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2016-01-29 | E-GEOD-77330 | biostudies-arrayexpress

Expression data from control (F/F) and Snai1 knockout (KO) embryoid body (EB) and teratoma ™

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2013-12-10 | E-GEOD-53106 | biostudies-arrayexpress

Transcription profiling of mouse Pooh cell line (POU5F1/Oct-4/3 overexpressing human)

Project description:POU5F1 (more commonly known as Oct-4/3) is one of the stem cell markers and affects direction of differentiation in embryonic stem cells. To investigate whether cells of mesenchymal origin acquire embryonic phenotype, we generated a human cell line of mesodermal origin with overexpression of the chimeric POU5F1 gene with physiological co-activator EWS, which is driven by the potent EWS promoter by translocation. The cell line termed Pooh (POU5F1/Oct-4/3 overexpressing human) cells expressed embryonic stem cell genes such as Nanog and also non-translocated POU5F1, lost mesenchymal phenotypes, and exhibited embryonal stem cell-like alveolar structure when implanted into the subcutaneous tissue of immunodeficient mice. Hierarchical analysis by microchip analysis and cell surface analysis revealed that Pooh cells are subcategorized into the group of human embryonic stem cells and embryonal carcinoma cells. These results imply that cells of mesenchymal origin can partially be traced back to cells to embryonic phenotype by the POU5F1 gene in collaboration with the potent cis-regulatory element and the fused co-activator. Experiment Overall Design: Pooh cells was generated from primary or first passage cells of pelvic tumor (Yamaguchi et al., Genes Chromosomes Cancer. 2005 Jun;43(2):217-22), and cultured in tissue culture dishes (100 mm, Becton Dickinson) in the G031101 medium (Med Shirotori, Tokyo). All cultures were maintained at 37Â°C in a humidified atmosphere containing 95% air and 5% CO2.

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