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