Metabolomics,Unknown,Transcriptomics,Genomics,Proteomics

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Gene expression profiles of C57BL/6 murine hepatocytes, murine embryonic stem cells (D3) and murine hepatocyte-reprogrammed induced Tissue Stem cells (iTSCs)

ABSTRACT: We have recently developed a new generation of induced stem cells that we have called “induced Tissue Stem cells” (iTSCs) which are generated by using low doses of non-integrative vectors encoding several embryonic reprogramming factors permitting to de-differentiate the cells without passing through a pluripotent state. ITSCs are thus multipotent stem cells rather than pluripotent stem cells. Using this technology we have produced endodermic iTSCs by de-differentiation of murine hepatocytes and shown that they are able to differentiate in vitro and in vivo into tissues restricted to endodermic layers. We report the gene expression profiles of two subclones of murine hepatocytes-reprogrammed iTSCs and of hepatocytes parental cells and murine embryonic stem cells (D3). Adherent liver cultures were established from 6 to 8 week old mice and one million of hepatocytes were infected with adenovirus encoding Oct4, Sox2, cMyc and Klf4. After 5 days of infections, cells were manually pooled and plated on mitomycin C-arrested Mouse Embryonic fibroblasts (MEF) in daily changed ES cell culture medium. In this manner, more than 50 colonies have been isolated and for two of them (clones 3 and 6), single cells were added in a 96-well cell culture plate for subcloning. Two subclones ; 6.6 and 3.8 were evaluated by microarray. The transcriptome was analyzed with Affymetrix microarray technology (GeneChip ® Mouse Gene 2.0 ST Array) and the data were normalized by the RMA method and annotated with Partek GS and R Bioconductor softwares.

ORGANISM(S): Mus musculus

SUBMITTER: christophe desterke

PROVIDER: E-GEOD-51782 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Project description:Chavez2009 - a core regulatory network of OCT4 in human embryonic stem cells A core OCT4-regulated network has been identified as a test case, to analyase stem cell characteristics and cellular differentiation. This model is described in the article: In silico identification of a core regulatory network of OCT4 in human embryonic stem cells using an integrated approach. Chavez L, Bais AS, Vingron M, Lehrach H, Adjaye J, Herwig R BMC Genomics, 2009, 10:314 Abstract: BACKGROUND: The transcription factor OCT4 is highly expressed in pluripotent embryonic stem cells which are derived from the inner cell mass of mammalian blastocysts. Pluripotency and self renewal are controlled by a transcription regulatory network governed by the transcription factors OCT4, SOX2 and NANOG. Recent studies on reprogramming somatic cells to induced pluripotent stem cells highlight OCT4 as a key regulator of pluripotency. RESULTS: We have carried out an integrated analysis of high-throughput data (ChIP-on-chip and RNAi experiments along with promoter sequence analysis of putative target genes) and identified a core OCT4 regulatory network in human embryonic stem cells consisting of 33 target genes. Enrichment analysis with these target genes revealed that this integrative analysis increases the functional information content by factors of 1.3 - 4.7 compared to the individual studies. In order to identify potential regulatory co-factors of OCT4, we performed a de novo motif analysis. In addition to known validated OCT4 motifs we obtained binding sites similar to motifs recognized by further regulators of pluripotency and development; e.g. the heterodimer of the transcription factors C-MYC and MAX, a prerequisite for C-MYC transcriptional activity that leads to cell growth and proliferation. CONCLUSION: Our analysis shows how heterogeneous functional information can be integrated in order to reconstruct gene regulatory networks. As a test case we identified a core OCT4-regulated network that is important for the analysis of stem cell characteristics and cellular differentiation. Functional information is largely enriched using different experimental results. The de novo motif discovery identified well-known regulators closely connected to the OCT4 network as well as potential new regulators of pluripotency and differentiation. These results provide the basis for further targeted functional studies. This model is hosted on BioModels Database and identified by: MODEL1305010000 . To cite BioModels Database, please use: BioModels Database: An enhanced, curated and annotated resource for published quantitative kinetic models . To the extent possible under law, all copyright and related or neighbouring rights to this encoded model have been dedicated to the public domain worldwide. Please refer to CC0 Public Domain Dedication for more information.

Dataset Information

Gene expression profiles of C57BL/6 murine hepatocytes, murine embryonic stem cells (D3) and murine hepatocyte-reprogrammed induced Tissue Stem cells (iTSCs)

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