Development of primary human nasal epithelial cell cultures for the study of cystic fibrosis pathophysiology

Francine de Courcey, Alexander V. Zholos, Hazel Atherton-Watson, Mark Thomas Shaw Williams, Paul Canning, Henry L. Danahay, J. Stuart Elborn, Madeleine Ennis

Research output: Contribution to journalArticle

23 Citations (Scopus)

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.

Original languageEnglish
Pages (from-to)C1173-c1179
JournalAmerican Journal of Physiology - Cell Physiology
Volume303
Issue number11
DOIs
Publication statusPublished - Dec 2012

Fingerprint Dive into the research topics of 'Development of primary human nasal epithelial cell cultures for the study of cystic fibrosis pathophysiology'. Together they form a unique fingerprint.

  • Cite this