title: Primary bronchial epithelial cell culture from explanted cystic fibrosis lungs.
abstract: Lung disease is responsible for more than 95% of morbidity and mortality in cystic fibrosis. The exact pathogenesis of cystic fibrosis lung disease remains poorly understood. Experimental models are therefore vital for use in research. Animal models and immortalized cell lines both have inherent limitations. Explanted lungs removed from people with cystic fibrosis at the time of transplantation represent a potentially valuable but technically and logistically challenging source of primary cystic fibrosis bronchial epithelial cells. In this study, pieces of segmental bronchus from explanted lungs were treated with patient-specific antimicrobials prior to isolation of bronchial epithelial cells. Cultured cells were characterized by their morphology under light microscopy, cytokeratin and hematoxylin-eosin staining, and electrophysiological profile. Primary bronchial epithelial cells were successfully cultured from 15 of 22 patients attempted. The cells exhibited typical epithelial morphology, staining for cytokeratin, lack of responsiveness to forskolin treatment, and remained viable after storage in liquid nitrogen. Seven unsuccessful cultures failed due to early infection with bacteria known to colonize the airways pretransplant. The results show that primary bronchial epithelial cell culture is possible from explanted cystic fibrosis lungs. This provides an important cellular model to elucidate the pathogenic mechanisms in cystic fibrosis lung disease and to investigate potential therapeutic targets.
authors.affiliation: Institute of Cellular Medicine, Newcastle University, Newcastle upon Tyne, United Kingdom. m.j.brodlie@ncl.ac.uk
authors.forename: Malcolm
authors.initials: M
authors.lastname: Brodlie
authors.forename: Michael C
authors.initials: MC
authors.lastname: McKean
authors.forename: Gail E
authors.initials: GE
authors.lastname: Johnson
authors.forename: John D
authors.initials: JD
authors.lastname: Perry
authors.forename: Audrey
authors.initials: A
authors.lastname: Nicholson
authors.forename: Bernard
authors.initials: B
authors.lastname: Verdon
authors.forename: Michael A
authors.initials: MA
authors.lastname: Gray
authors.forename: John H
authors.initials: JH
authors.lastname: Dark
authors.forename: Jeffrey P
authors.initials: JP
authors.lastname: Pearson
authors.forename: Andrew J
authors.initials: AJ
authors.lastname: Fisher
authors.forename: Paul A
authors.initials: PA
authors.lastname: Corris
authors.forename: James
authors.initials: J
authors.lastname: Lordan
authors.forename: Christopher
authors.initials: C
authors.lastname: Ward
mesh_terms: D000328:Adult; D000890:Anti-Infective Agents; D018929:Cell Culture Techniques; D002478:Cells, Cultured; D003550:Cystic Fibrosis / Q000473:pathology*; D004847:Epithelial Cells / Q000166:cytology*; D006801:Humans; D008168:Lung / Q000473:pathology*; D008875:Middle Aged; D055815:Young Adult
chemical_list: D000890:Anti-Infective Agents