E-GEOD-51782 - Gene expression profiles of C57BL/6 murine hepatocytes, murine embryonic stem cells (D3) and murine hepatocyte-reprogrammed induced Tissue Stem cells (iTSCs)
Released on 29 October 2013, last updated on 4 November 2013
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.
transcription profiling by array
christophe desterke <firstname.lastname@example.org>, Annelise Bennaceur-Griscelli, Christophe Desterke, Frank Griscelli