Fibroblasts and transforming growth factor β induce organization and differentiation of T84 human epithelial cells

Abstract

Background and Aims: The gut epithelium in the cryptvillus axis represents a continuous developmental system in which the role of fibroblast- epithelial interactions is obvious. The aim of this study was to establish an in vitro method whereby fibroblast-guided differentiation of crypt-like gut epithelial cells can be studied. Methods: intestinal epithelial cells (T84 and HT-29) were cultured within type I collagen gel together with fibroblasts without cell-to-cell contact. T84 cells were also grown in the presence of transforming growth factor β and hepatocyte growth factor. The gels were studied using light and electron microscopy and histochemical and immunohistochemical methods. Results: The epithelial cells formed unorganized cell clusters within the gels, but when given fibroblast support, 76% of the T84 cell colonies (not HT-29) organized into luminal formations, and basement membranes including laminin were well deposited. The cells in the columnar single cell-layer luminal formations (49% of all colonies) were differentiated, showing microvilli, up-regulated alkaline phosphatase brush border activity, and mucin profiles typical for small intestine. This fibroblast-induced organization and differentiation was induced by transforming growth factor β. Conclusions: Crypt-like T84 epithelial cells are able to differentiate when grown three-dimensionally together with fibroblasts or transforming growth factor β. This method may be used for mesenchymal-epithelial cell cross-talk studies.