Absorptive and mucus-secreting subclones isolated from a multipotent intestinal cell line (HT-29) provide new models for cell polarity and terminal differentiation.
A clone HT29-18 has been isolated from the parent cell line HT-29, which derived from a human colon adenocarcinoma (Fogh, J., and G. Trempe, 1975, Human Tumor Cells in Vitro, J. Fogh, editor, Plenum Publishing Corp., New York, 115-141). This clone is able to differentiate as the parent cell line does. Differentiation occurs when glucose is replaced by galactose in the culture medium (Pinto, M., M.D. Appay, P. Simon-Assman, G. Chevalier, N. Dracopoli, J. Fogh, and A. Zweibaum, 1982, Biol. Cell., 44:193-196). We demonstrate here that the differentiated cloned population HT29-18/gal is heterogenous: although 90% of the cells show morphological characteristics of "absorptive cells", only 20-30% of them display sucrase-isomaltase in their apical microvillar membranes. About 10% of the entire cell population consists of cells containing mucous granules similar to intestinal goblet cells. We have isolated two subclones, HT29-18-C1 and HT29-18-N2, from the differentiated HT29-18/gal cells. HT29-18-C1 cells show morphological characteristics of polarized absorptive cells, when growing either in glucose- or in galactose-containing media, but the sucrase-isomaltase is not expressed in the cells grown in glucose-containing medium. The clone HT29-18-N2 is also polarized in both culture conditions and is similar to globlet cells in vivo. It grows as a monolayer, exhibits tight junctions, and contains numerous mucous granules whose exocytosis can be triggered by carbachol, a parasympathomimetic drug. We conclude that the clone HT29-18 first isolated was a multipotent cell population from which we isolated several subclones that differentiate either as absorptive (HT29-18-C1) or as mucous (HT29-18-N2) cells. In contrast to the parent HT-29 cell line, the subclones retain most of their differentiated properties in glucose-containing medium.
D000230:Adenocarcinoma / Q000473:pathology*; D002217:Carbachol / Q000494:pharmacology; D002454:Cell Differentiation / Q000187:drug effects; D002460:Cell Line; D002999:Clone Cells / Q000378:metabolism / Q000648:ultrastructure*; D003110:Colonic Neoplasms / Q000473:pathology*; D003594:Cytoplasmic Granules / Q000648:ultrastructure; D005089:Exocytosis / Q000187:drug effects; D005690:Galactose / Q000494:pharmacology; D006801:Humans; D007408:Intestinal Absorption; D008565:Membrane Proteins / Q000032:analysis; D008871:Microvilli / Q000201:enzymology / Q000648:ultrastructure; D008954:Models, Biological; D009093:Mucus / Q000378:metabolism; D009834:Oligo-1,6-Glucosidase / Q000032:analysis; D013393:Sucrase / Q000032:analysis
D008565:Membrane Proteins; D002217:Carbachol; D009834:Oligo-1,6-Glucosidase; D013393:Sucrase; D005690:Galactose