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    Small molecule and RNAi induced phenotype transition of expanded and primary colonic epithelial cells (2015)

    Art
    Poster
    Autoren
    Sharbati, Jutta (WE 3)
    Hanisch, Carlos (WE 3)
    Pieper, Robert (WE 4)
    Einspanier, Ralf (WE 3)
    Sharbati, Soroush (WE 3)
    Forschungsprojekt
    Verification of GMO risk assessment elements and review and communication of evidence collected on the biosafety of GMO (GRACE)
    Kongress
    Non-coding Genome EMBL
    Heidelberg, 18. – 21.10.2015
    Quelle
    Scientific reports; 5(12681) — S. 1–11
    ISSN: 2045-2322
    Sprache
    Englisch
    Verweise
    URL (Volltext): http://www.nature.com/articles/srep12681
    DOI: 10.1038/srep12681
    Kontakt
    Institut für Veterinär-Biochemie

    Oertzenweg 19 b
    14163 Berlin
    +49 30 838 62225
    biochemie@vetmed.fu-berlin.de

    Abstract / Zusammenfassung

    Recent progress in mammalian intestinal epithelial cell culture led to novel concepts of tissue modeling. Especially the development of phenotypically stable cell lines from individual animals enables an investigation of distinct intestinal loci and disease states. We here report primary and prolonged culture of normal porcine epithelial cells from colon for cell line development. In addition, a novel primary three-dimensional intestinal culture system is presented, which generated organoids composed of a highly polarized epithelial layer lining a core of subepithelial tissue. Cellular characterization of monolayer cell lines revealed epithelial identity and pointed to a proliferative crypt cell phenotype. We evaluated both RNAi and chemical approaches to induce epithelial differentiation in generated cell lines by targeting promoters of epithelial to mesenchymal transition (EMT). By in silico prediction and ectopic expression, miR-147b was proven to be a potent trigger of intestinal epithelial cell differentiation. Our results outline an approach to generate phenotypically stable cell lines expanded from primary colonic epithelial cultures and demonstrate the relevance of miR-147b and chemical inhibitors for promoting epithelial differentiation features.