title: IEC-18, a nontransformed small intestinal cell line for studying epithelial permeability.
abstract: Small intestinal epithelium is leaky and allows permeation of hydrophilic molecules of various sizes. Passively absorbed hydrophilic permeability probes have been shown to permeate across intestinal epithelium mainly through the paracellular pathways. In this study we introduce microporous filter-grown IEC-18 epithelial cells, a nontransformed small intestinal cell line, as a in vitro model of intestinal epithelium for the study of epithelial permeability. IEC-18 cells, originally derived from native rat ileal crypts, form confluent epithelium when grown on hydrated collagen-coated Millicell-CM permeable inserts (Millipore Corp., Bedford, Mass.). With scanning and transmission electron microscopy, the presence of tight junctions and desmosomes between cells and the development of microvilli at the apical surface were confirmed. Immunofluorescent labeling of ZO-1 proteins and desmoplakins verified the presence of tight-junctional proteins (ZO-1) and desmosomes in the intercellular junctions of confluent IEC-18 epithelium. The net electrical resistance of IEC-18 epithelium (28 omega-cm2) was similar to resistance values obtained from small intestinal tissue with (50 to 100 omega-cm2) or without (20 to 45 omega-cm2) muscularis and serosal layers. Assessment of mannitol and dextran permeation revealed early "maturation" of paracellular pathway, with increasing restriction of permeation to both probes through day 4. Resistance across IEC-18 epithelium also reached plateau levels between 4 and 7 days. Permeability studies with various probes indicate that cross-sectional diameter rather than molecular weight of the probe is the important determinant of permeation rate. IEC-18 epithelium selectively restricted the permeation of probes proportional to probe size; permeation of larger probes such as albumin was negligible. We conclude that cultured IEC-18 epithelial cells, because of their native crypt origin, similarity in resistance to small intestinal epithelia, retention of ability to differentiate into villus-like enterocytes, and permeability characteristics, are a useful model of intestinal epithelium for the study of permeability and paracellular transport.
authors.affiliation: Department of Medicine, University of California, Irvine, Long Beach.
authors.forename: T Y
authors.initials: TY
authors.lastname: Ma
authors.forename: D
authors.initials: D
authors.lastname: Hollander
authors.forename: D
authors.initials: D
authors.lastname: Bhalla
authors.forename: H
authors.initials: H
authors.lastname: Nguyen
authors.forename: P
authors.initials: P
authors.lastname: Krugliak
mesh_terms: D000818:Animals; D002450:Cell Communication; D002460:Cell Line; D002462:Cell Membrane / Q000502:physiology / Q000648:ultrastructure; D002463:Cell Membrane Permeability; D003896:Desmosomes / Q000502:physiology / Q000648:ultrastructure; D004848:Epithelium / Q000502:physiology / Q000648:ultrastructure; D007365:Intercellular Junctions / Q000502:physiology / Q000648:ultrastructure; D007421:Intestine, Small / Q000502:physiology*; D008855:Microscopy, Electron, Scanning; D008871:Microvilli / Q000502:physiology / Q000648:ultrastructure; D051381:Rats