Migration of IEC-6 cells: a model for mucosal healing

Abstract

Cell migration is the principal force behind the early restitution of erosions of the mucosa of the gastrointestinal tract. Despite the importance of cell migration to healing, no attempts to study the process in culture have been reported. We have attempted to standardize conditions for migration and test the migration responses of the small intestinal epithelial crypt cell line IEC-6 in some experimental situations already well known in vivo. We found good correspondence between in culture and in vivo on the following points: 1) migration was independent of DNA synthesis; 2) DNA synthesis was not concentrated at the wound edge; and 3) inhibition of actin polymerization stopped migration altogether. In addition, the presence of an extracellular matrix maximized migration. Protein inhibitors with different modes of action inhibited cell migration to different degrees, not always commensurate with their inhibition of protein synthesis. Cell surface proteoglycans were important; hyaluronic acid had an effect, but the secretion of a migration-stimulating substance by wounded cells was equivocal. Significantly, alpha-difluoromethylornithine (DFMO), which inhibits ornithine decarboxylase and polyamine synthesis, almost totally prevented cell migration. Because DFMO also prevents healing of mucosal erosions in vivo, we believe that this model can be used, keeping in mind its spatial limitations, to study the process of cell migration involved in the early restitution of mucosal erosions.

ANG II and LPA induce Pyk2 tyrosine phosphorylation in intestinal epithelial cells: role of Ca2+, PKC, and Rho kinase

“…The IEC-18 and IEC-6 small intestinal cell lines have provided a useful model for studying these functions in culture (7,9,33,40,55). Although growth factor-induced proliferation and motility have been described in these cells, the associated intracellular signaling mechanisms are not well characterized.…”

“…INTESTINAL EPITHELIAL CELL FUNCTIONS, including growth, motility, differentiation, and transport, are regulated by a combination of environmental factors that include peptide signals, bioactive lipids, adhesion to the extracellular matrix, and cell wounding (2,7,13,33,47,55). The nontransformed IEC-18 and IEC-6 intestinal epithelial cell lines, derived from rat small intestinal crypt epithelium (40), have been widely used as a model of epithelial cell proliferation, migration, and differentiation (7,9,33,55).…”

“…The nontransformed IEC-18 and IEC-6 intestinal epithelial cell lines, derived from rat small intestinal crypt epithelium (40), have been widely used as a model of epithelial cell proliferation, migration, and differentiation (7,9,33,55). In the "artificial wound" model of epithelial restitution, IEC-6 cell migration is promoted by transforming growth factors-␣ and -␤ (TGF-␣ and -␤), epidermal growth factor (EGF), and lysophosphatidic acid (LPA).…”

Alanyl-Glutamine and Glutamine Supplementation Improves 5-Fluorouracil-Induced Intestinal Epithelium Damage In Vitro

“…Rat intestinal jejunal crypt cells (IEC-6, passages [20][21][22][23][24][25][26][27][28][29][30][31][32][33][34][35] were purchased from American Type Culture Collection (Rockville, MD) and were cultured at 37°C in a 5% CO 2 incubator. The maintenance cell media was Dulbecco's modified eagle media (DMEM; Gibco BRL, Grand Island, NY) supplemented with 5% fetal calf serum (FCS), 5 mg bovine insulin, 50 μg/ml penicillin/streptomycin (DMEM; Gibco BRL, Grand Island, NY), and a final concentration of 1 mM sodium pyruvate.…”

“…Afterwards, wells were washed using DMEM. Wells were digitally photographed after 6, 12, and 24 h, and the number of cells in the longest migration line was counted and averaged according to the well surface area (adapted from McCormack and Brito et al [28]). The digital pictures were captured and opened using Image Pro Plus v. 5.0 software (Media Cybernetics, Silver Spring, MD).…”