Unknown,Transcriptomics,Genomics,Proteomics

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Transcription profiling of mouse ES cells and EpiSCs

ABSTRACT: The application of human embryonic stem (ES) cells in medicine; and biology has an inherent reliance on understanding the starting; cell population. Human ES cells differ from mouse ES cells and the; specific embryonic origin of both cell types is unclear. Previous; work suggested that mouse ES cells could only be obtained from; the embryo before implantation in the uterus1â5. Here we show; that cell lines can be derived from the epiblast, a tissue of the postimplantation; embryo that generates the embryo proper. These; cells, which we refer to as EpiSCs (post-implantation epiblastderived; ES cells), express transcription factors known to regulate; pluripotency, maintain their genomic integrity, and robustly differentiate; into the major somatic cell types as well as primordial; germ cells. The EpiSC lines are distinct from mouse ES cells in; their epigenetic state and the signals controlling their differentiation. Furthermore, EpiSC and human ES cells share patterns of; gene expression and signalling responses that normally function; in the epiblast. These results show that epiblast cells can be maintained; as stable cell lines and interrogated to understand how; pluripotent cells generate distinct fates during early development. *Note: EpiSCs were previously referred to as post-ES cells Experiment Overall Design: 3 biological replicates of each cell type (EpiSC and mouse ES) were compared.

ORGANISM(S): Mus musculus

SUBMITTER: Paul Tesar

PROVIDER: E-GEOD-7866 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Publications

New cell lines from mouse epiblast share defining features with human embryonic stem cells.

Tesar Paul J PJ Chenoweth Josh G JG Brook Frances A FA Davies Timothy J TJ Evans Edward P EP Mack David L DL Gardner Richard L RL McKay Ronald D G RD

Nature 20070627 7150

The application of human embryonic stem (ES) cells in medicine and biology has an inherent reliance on understanding the starting cell population. Human ES cells differ from mouse ES cells and the specific embryonic origin of both cell types is unclear. Previous work suggested that mouse ES cells could only be obtained from the embryo before implantation in the uterus. Here we show that cell lines can be derived from the epiblast, a tissue of the post-implantation embryo that generates the embry ...[more]

PMID: 17597760

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Project description:Recently, (in vitro) pluripotent EpiSCs were derived from the post-implantation egg cylinder stage epiblasts of mouse and rat. These EpiSCs resemble and correspond very closely to the conventional human embryonic stem cells (hESCs) in the colony morphology and culture/signaling requirements for maintaining pluripotency, but exhibit a range of significant phenotypic and signaling response differences from the conventional mouse ES cells (mESCs). These observations strongly support the notion that EpiSCs and hESCs are intrinsically similar, and raise an attractive hypothesis: as mESCs and EpiSCs/hESCs represent two distinct pluripotency states: the mESC-like state representing the ICM of pre-implantation blastcyst and the EpiSC-like state representing the post-implantation epiblasts, whether the epiblast state (including conventional hESCs) can be converted back to the ICM state. Despite studies providing evidence that epiblast-like cells exist and transition back and forth within colony of conventional mESCs; mESCs and EpiSCs share substantial set of pluripotency transcriptional factors, including Oct4, Sox2 and Nanog; and mESCs are more stable in culture, in the present study we found that EpiSCs differentiated rapidly under mESC culture conditions and no âspontaneouslyâ converted mESC could be readily identified and isolated over serial passages at the population or clonal level. Remarkably, we found that blockage of the TGFÎ² pathway or inhibition of the H3K4 demethylase LSD1 with small molecule inhibitors induced dramatic morphological changes of EpiSCs towards mESC phenotypes with activation of some ICM-specific gene expression. However, full conversion of EpiSCs to a mESC-like state with competence to chimeric contribution can only be readily generated with a combination of inhibitors of LSD1 and ALK. These observations underscore a powerful and direct induction of reprogramming from the developmentally later-stage EpiSCs to a mESC-like stage by a synergy of signaling and direct epigenetic modulations. It also highlights a significant role for TGFÎ² pathway inhibition in promoting reprogramming to and sustaining true pluripotency, which further supports our recent studies in generating chimerism-competent rat pluripotent cells. Collectively, our studies provide a proof-of-concept demonstration that pluripotency-restricted EpiSCs can be readily converted to a mESC-like state in the absence of any genetic manipulation by precise pharmacological control of signaling pathways that distinguish the two pluripotency states and an epigenetic target simultaneously, and offer a convenient experimental system to further study the mechanism. Such method and concept may also provide an avenue for generating a new type of mESC-like human pluripotent cell. Global gene-expression analyses of the parnate/mAMFGi cells

Dataset Information

Transcription profiling of mouse ES cells and EpiSCs

Publications

New cell lines from mouse epiblast share defining features with human embryonic stem cells.

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