title: Development of primary human nasal epithelial cell cultures for the study of cystic fibrosis pathophysiology.
abstract: Cultured primary epithelial cells are used to examine inflammation in cystic fibrosis (CF). We describe a new human model system using cultured nasal brushings. Nasal brushings were obtained from 16 F508del homozygous patients and 11 healthy controls. Cells were resuspended in airway epithelial growth medium and seeded onto collagen-coated flasks and membranes for use in patch-clamp, ion transport, and mediator release assays. Viable cultures were obtained with a 75% success rate from subjects with CF and 100% from control subjects. Amiloride-sensitive epithelial Na channel current of similar size was present in both cell types while forskolin-activated CF transmembrane conductance regulator current was lacking in CF cells. In Ussing chambers, cells from CF patients responded to UTP but not to forskolin. Spontaneous and cytomix-stimulated IL-8 release was similar (stimulated 29,448 ± 9,025 pg/ml; control 16,336 ± 3,308 pg/ml CF; means ± SE). Thus nasal epithelial cells from patients with CF can be grown from nasal brushings and used in electrophysiological and mediator release studies in CF research.
authors.affiliation: Centre for Infection and Immunity, Queen’s University Belfast, Health Sciences Building, Belfast, UK.
authors.forename: F
authors.initials: F
authors.lastname: de Courcey
authors.forename: A V
authors.initials: AV
authors.lastname: Zholos
authors.forename: H
authors.initials: H
authors.lastname: Atherton-Watson
authors.forename: M T S
authors.initials: MT
authors.lastname: Williams
authors.forename: P
authors.initials: P
authors.lastname: Canning
authors.forename: H L
authors.initials: HL
authors.lastname: Danahay
authors.forename: J S
authors.initials: JS
authors.lastname: Elborn
authors.forename: M
authors.initials: M
authors.lastname: Ennis
mesh_terms: D000328:Adult; D000584:Amiloride / Q000494:pharmacology; D002478:Cells, Cultured; D005576:Colforsin / Q000494:pharmacology; D003550:Cystic Fibrosis / Q000188:drug therapy / Q000503:physiopathology*; D019005:Cystic Fibrosis Transmembrane Conductance Regulator / Q000235:genetics / Q000502:physiology; D062686:Epithelial Sodium Channel Blockers / Q000494:pharmacology; D005260:Female; D006801:Humans; D053583:Interleukin-1beta / Q000494:pharmacology; D016209:Interleukin-8 / Q000378:metabolism; D008070:Lipopolysaccharides / Q000494:pharmacology; D008297:Male; D017817:Nasal Lavage Fluid; D009297:Nasal Mucosa / Q000187:drug effects / Q000503:physiopathology*; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology; D014544:Uridine Triphosphate / Q000494:pharmacology; D055815:Young Adult
chemical_list: C508606:CXCL8 protein, human; D062686:Epithelial Sodium Channel Blockers; C508585:IL1B protein, human; D053583:Interleukin-1beta; D016209:Interleukin-8; D008070:Lipopolysaccharides; D014409:Tumor Necrosis Factor-alpha; D019005:Cystic Fibrosis Transmembrane Conductance Regulator; D005576:Colforsin; D000584:Amiloride; D014544:Uridine Triphosphate