Published ahead of print on September 22, 2005, doi:10.1165/rcmb.2005-0161OC
© 2006 American Thoracic Society DOI: 10.1165/rcmb.2005-0161OC Long-Term Cultures of Polarized Airway Epithelial Cells from Patients with Cystic FibrosisLaboratory of Clinical Investigation III, Department of Pediatrics, Division of Pulmonary Medicine, and Clinic of Oto-Rhino-Laryngology, Geneva University Hospitals, Geneva; Department of Pathology, Geneva Medical School, Geneva, Switzerland Correspondence and requests for reprints should be addressed to Marc Chanson, Ph.D., Laboratory of Clinical Investigation III, Department of Pediatrics, HUG-P.O. Box 14, Micheli-du-Crest, 24, 1211 Geneva 14, Switzerland. E-mail: Marc.Chanson{at}hcuge.ch 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.
Key Words: airway epithelial cells • cystic fibrosis • epithelial–mesenchymal interactions • human cell model • long-term cell differentiation This article has been cited by other articles:
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