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Expression data from undifferentiated human ES cell line, khES3, grown using complete or methionine deprived media and ES cell derived endoderm

ABSTRACT: In undifferentiated human ES cells, 5hr Met deprivation (delta Met) led to decreased proliferation, and prolonged 24hr Met deprivation resulted in G0-G1 phase cell cycle arrest, which then led to apoptosis. We used microarrays to detail the molecular mechanism of cell-death caused by Met deprivation in undifferentiated ES cells and checked the difference of Met cycle relate gene expression between undifferentiated human ES cells and ES cell derived endoderm. Cells were harvested at undifferentiated or differentiated stages for RNA extraction and hybridization on Affymetrix microarrays. We performed gene expression profiles of undifferentiated khES3 cells cultured in complete (compl) or Met-deprived conditions (deltaMet) for 5 or 24 hr. And 0hr untreated cells and endoderm cells were used as a comparison

ORGANISM(S): Homo sapiens

SUBMITTER: nobuaki shiraki

PROVIDER: E-GEOD-55283 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Expression data from undifferentiated human ES cell line, khES3, grown using complete or methionine deprived media and ES cell derived endoderm

Project description:In undifferentiated human ES cells, 5hr Met deprivation (delta Met) led to decreased proliferation, and prolonged 24hr Met deprivation resulted in G0-G1 phase cell cycle arrest, which then led to apoptosis. We used microarrays to detail the molecular mechanism of cell-death caused by Met deprivation in undifferentiated ES cells and checked the difference of Met cycle relate gene expression between undifferentiated human ES cells and ES cell derived endoderm.

2014-04-17 | GSE55283 | GEO

Expression data from undifferentiated human iPS cell line, 201B7

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Expression data from undifferentiated human ES cell line, khES3 grown using complete, leucine or lysine deprived media

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Expression data from undifferentiated human ES cell line, khES3 grown using complete, leucine or lysine deprived media

Project description:In undifferentiated human ES cells, 48hr Leucine deprivation (delta Leu) or Lysine deprivation (delta Lys) led to apoptosis. We used microarrays to detail the mollecular mechanism of cell-death caused by Leu, Lys deprivation in undifferentiated ES cells. Cells were harvested at undifferentiated stages for RNA extraction and hybridization on Affymetrix microarrays. We performed gene expression profiles of undifferentiated khES3 cells cultured in complete (comple),Leu or Lys deprivation for 5 hr.

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Gene expression profiling of Nodal versus Activin-derived endoderm from mouse ES cells

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Gene expression profiling of Nodal versus Activin-derived endoderm from mouse ES cells

Project description:To explore the molecular basis of functional differences observed between Nodal versus Activin-derived endoderm, we compared their respective gene expression profiles. Sox17-GFP mouse ES cells were differentiated in the presence of Nodal or Activin for 7 days, after which GFP(+) cells were purified by FACs. Undifferentiated ES cells were also included for comparison as a control. Results indicate that the two endoderm populations are nearly identical at the level of global transcription. Subtle differences suggest a difference in the degree of endoderm progression. Total RNA obtained from SOX17-GFP (+) mouse ES cells, Sox17-GFP treated with Nodal, Sox17-GFP treated with Activin. Two replicates for each sample; Sample data table contains average values of two replicates. The non_normalized.txt file contains individual signal values for each replicate.

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RNA-seq of human induced pluripotent cells treated with short-term Met deprival

Project description:Human iPSCs require high amounts of methionine (Met). Met deprivation results in a rapid decrease in intracellular S-adenosyl-methionine (SAM), poising human iPSCs for differentiation and leading to apoptosis of undifferentiated cells. Met deprivation triggers a rapid metabolite change, including SAM, followed by reversible epigenetic modification. We show here that short-term Met deprivation impairs pluripotency network through epigenetic modification. The trimethylation of lysine 4 on histone H3 (H3K4me3) was dramatically affected compared to other histone modifications. Transcription start site (TSS) region of key pluripotent genes such as NANOG and OCT3/4 are specifically impacted upon short-term Met deprivation. Gene expression levels of these genes decreased, correlating to the loss of H3K4me3 marks. Upon differentiation, Met deprivation triggers a loss of the H3K27me3 in many mesendodermal genes, thereby switching from a bivalent to monovalent (H3K4me3) state. We concluded that Met metabolism maintains the pluripotent network with histone marks, and their loss potentiates differentiation.

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Expression data from ES cell-derived motoneuron

Project description:Neural formation from ES cells provides a novel system for studying axonogenesis in projection neurons. We used microarrays to clarify the global gene expression patterns of ES-derived motoneurons and compared with the parental undifferentiated ES cells.

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GRO-seq in mouse Epiblast-primed, Primitive endoderm-primed, and differentiated endoderm cells

Project description:The identification of changes in transcriptional regulation during priming and differentiation of embryonic stem (ES) cells towards the endoderm lineage. Specific populations of ES cells, either primed or committed to endoderm, were isolated and subjected to global nuclear run on sequencing (GRO-Seq). The hHex-Venus (HV) reporter ES cell line, HVJu5.1 (Canham et al., 2010) was used to isolate HV- and HV+ ES cells. Primed ES cells were identified based on the expression of the HV marker in addition to the cell surface marker of undifferentiated ES cells, SSEA-1, (the lower and upper 25% of SSEA-1+, HV expressing cells). When challenged to differentiate, HV- ES cells are primed towards an epiblast fate, while HV+ ES cells are primed towards primitive endoderm. However, these populations are considered primed, rather than committed, as they will readily interconvert when re-introduced into standard ES cell culture conditions. ES cells were grown in self-renewing conditions (GMEM, LIF, 10% FCS, plated on gelatin coated dishes). Endoderm was obtained by differentiating ES cells in medium without the cytokine LIF for 5 days. The HV+, SSEA-1- differentiated fraction was then sorted and represents an early stage in endoderm differentiation.

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