title: Long-term cultures of polarized airway epithelial cells from patients with cystic fibrosis.
abstract: The poor ability of respiratory epithelial cells to proliferate and differentiate in vitro into a pseudostratified mucociliated epithelium limits the general use of primary airway epithelial cell (AEC) cultures generated from patients with rare diseases, such as cystic fibrosis (CF). Here, we describe a procedure to amplify AEC isolated from nasal polyps and generate long-term cultures of the respiratory epithelium. AEC were seeded onto microporous permeable supports that carried on their undersurface a preformed feeder layer of primary human airway fibroblasts. The use of fibroblast feeder layers strongly stimulated the proliferation of epithelial cells, allowing the expansion of the cell pool with successive passages. AEC at increasing passage were seeded onto supports undercoated with airway fibroblasts and exposed to air. Either freshly isolated or amplified AEC could differentiate into a pseudostratified mucociliated epithelium for at least 10 mo. Thus, CF epithelia cultures showed elevated Na+ transport, drastic hyperabsorption of surface liquid, and absence of cAMP-induced Cl- secretion as compared with non-CF cultures. They were also characterized by thick apical secretion that hampered the movement of cell surface debris by cilia. However, CF respiratory epithelia did not show increased production of mucins or IL-8. The method described here is now routinely used in our laboratory to establish long-term cultures of well differentiated respiratory epithelia from human airway biopsies.
authors.affiliation: Laboratory of Clinical Investigation III, Department of Pediatrics, HUG, Geneva 14, Switzerland.
authors.forename: Ludovic
authors.initials: L
authors.lastname: Wiszniewski
authors.forename: Lan
authors.initials: L
authors.lastname: Jornot
authors.forename: Tecla
authors.initials: T
authors.lastname: Dudez
authors.forename: Alessandra
authors.initials: A
authors.lastname: Pagano
authors.forename: Thierry
authors.initials: T
authors.lastname: Rochat
authors.forename: Jean Silvain
authors.initials: JS
authors.lastname: Lacroix
authors.forename: Susanne
authors.initials: S
authors.lastname: Suter
authors.forename: Marc
authors.initials: M
authors.lastname: Chanson
mesh_terms: D001692:Biological Transport / Q000502:physiology; D018929:Cell Culture Techniques; D002454:Cell Differentiation / Q000502:physiology; D016764:Cell Polarity; D048430:Cell Shape; D002478:Cells, Cultured; D003550:Cystic Fibrosis / Q000503:physiopathology*; D019005:Cystic Fibrosis Transmembrane Conductance Regulator / Q000378:metabolism; D004594:Electrophysiology; D004847:Epithelial Cells / Q000166:cytology / Q000378:metabolism; D005347:Fibroblasts / Q000166:cytology / Q000378:metabolism; D006801:Humans; D016209:Interleukin-8 / Q000378:metabolism; D009077:Mucins / Q000378:metabolism; D020545:Respiratory Mucosa / Q000166:cytology*; D013234:Stem Cells
chemical_list: C505032:CFTR protein, human; D016209:Interleukin-8; D009077:Mucins; D019005:Cystic Fibrosis Transmembrane Conductance Regulator