title: The NCI-N87 cell line as a gastric epithelial barrier model for drug permeability assay.
abstract: The objective of this study was to evaluate the human NCI-N87 cell line as a model for gastric permeability drug studies under pH conditions of the stomach. The optimal conditions that led NCI-N87 cells to form a typical differentiated gastric epithelial barrier were a seeding density of 2.5 × 10⁵ cells/cm² on porous inserts and growth in serum-complemented RPMI-1640 medium until 18-27 days post-confluency. The resulting cell monolayers showed moderately high transepithelial electrical resistance (TEER) values of about 500 Ω cm², cells of polygonal morphology expressing E-cadherin and ZO-1 proteins at their contact surfaces, and production of mucus clusters. The monolayers withstood apical pH of 7.4 down to 3.0 with the basal pH fixed at 7.4. The apparent permeability coefficients (P(app)) of model compounds were evaluated in the apical-to-basolateral and basolateral-to-apical directions under different pH gradients. The monolayers were impermeable to the integrity marker Lucifer Yellow (low P(app) of 0.3-1.1 × 10⁻⁶ cm/s). The furosemide P(app) (0.4-1.5 × 10⁻⁵ cm/s) were slightly dependent on pH but remained moderate. The caffeine P(app) (4.2-5.0 × 10⁻⁵ cm/s) were higher and insensitive to pH changes. The NCI-N87 cell line provides a useful in vitro tool to assess gastric drug permeability and absorption under physiologic conditions prevailing in the human stomach.
authors.affiliation: Department of Chemistry-Biochemistry and Centre Pharmaqam, Université du Québec à Montréal, CP 8888, Succ. Centre-ville, Montréal, Québec, Canada.
authors.forename: Marc
authors.initials: M
authors.lastname: Lemieux
authors.forename: Frédéric
authors.initials: F
authors.lastname: Bouchard
authors.forename: Patrick
authors.initials: P
authors.lastname: Gosselin
authors.forename: Joanne
authors.initials: J
authors.lastname: Paquin
authors.forename: Mircea Alexandru
authors.initials: MA
authors.lastname: Mateescu
mesh_terms: D000042:Absorption; D015820:Cadherins / Q000378:metabolism; D002110:Caffeine / Q000493:pharmacokinetics; D002460:Cell Line; D004341:Drug Evaluation / Q000379:methods*; D017097:Electric Impedance; D005665:Furosemide / Q000493:pharmacokinetics; D005753:Gastric Mucosa / Q000378:metabolism*; D006801:Humans; D006863:Hydrogen-Ion Concentration; D008565:Membrane Proteins / Q000378:metabolism; D008954:Models, Biological; D010599:Pharmacokinetics; D010750:Phosphoproteins / Q000378:metabolism; D062826:Zonula Occludens-1 Protein
chemical_list: D015820:Cadherins; D008565:Membrane Proteins; D010750:Phosphoproteins; C569042:TJP1 protein, human; D062826:Zonula Occludens-1 Protein; D002110:Caffeine; D005665:Furosemide