Phenotypic characterization of an intestinal subepithelial myofibroblast cell line

Abstract

Subepithelial myofibroblasts are located at the interface between the epithelium and lamina propria in most mucosal tissues. Their biological functions are largely unknown because a long-term cell culture model for these cells has not been available. In this report, we define the phenotypic properties of a human colonic cell line (18Co) that exhibits most of the known characteristics of intestinal subepithelial myofibroblasts in situ. These characteristics include 1) a cell shape that can be reversibly interconverted between a flattened discoid and stellate morphology, 2) intracellular organelles reminiscent of myofibroblasts and smooth muscle cells in situ, 3) expression of alpha-smooth muscle actin, 4) plasma membrane receptors for endothelins and natriuretic peptides, and 5) regulation of epithelial sensitivity to calcium-dependent secretagogues by paracrine secretion of prostaglandins. 18Co cells provide an exploitable model to begin defining the physiological and pathophysiological functions of intestinal subepithelial myofibroblasts at the molecular level.

Regulation of COX-2 expression in human intestinal myofibroblasts: mechanisms of IL-1-mediated induction

“…The human colonic subepithelial myofibroblast cell line, 18Co was obtained from the American Type Culture Collection (CRL-1459) and maintained as described previously (56). Culture media, supplements, and subculturing reagents were purchased from Sigma.…”

“…As such, the COX-2 gene is subject to regulation by numerous signaling pathways, and the relative contribution of each depends upon the stimulus, the cellular environment, and the particular cell type. We have characterized a myofibroblast cell line, 18Co, derived from mucosal biopsy of human neonatal colon, which exhibits the phenotypic features of primary ISEMF cultures and ISEMFs in situ (56). Herein, using IL-1 as a model stimulus, we report the first detailed analysis of signaling pathways important for COX-2 expression in human intestinal myofibroblasts and how these pathways interact with each other.…”

Subepithelial Myofibroblasts are Novel Nonprofessional APCs in the Human Colonic Mucosa

“…A commercially available human CMF primary isolate, 18Co, was also used as a representative CMF (45). Phenotypic characterization of cultured CMFs and 18Co cells has demonstrated that they are uniformly positive for ␣-SMA and CD90, but negative for desmin (Fig.…”

PAR1-dependent and independent increases in COX-2 and PGE2 in human colonic myofibroblasts stimulated by thrombin

“…MYOFIBROBLASTS ARE LOCATED in the lamina propria of the intestine in close proximity to epithelial cells and have ultrastructural features of both smooth muscle cells and fibroblasts (38). They are paracrine cells that regulate a number of fundamental biological processes in the intestine, including epithelial chloride secretion (11), which, in turn, controls the secretion of water into the intestinal lumen.…”