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Title

Long-term cultures of polarized airway epithelial cells from patients with cystic fibrosis

Authors
Published in American journal of respiratory cell and molecular biology. 2006, vol. 34, no. 1, p. 39-48
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.
Keywords Biological Transport/physiologyCell Culture TechniquesCell Differentiation/physiologyCell PolarityCell ShapeCells, CulturedCystic Fibrosis/physiopathologyCystic Fibrosis Transmembrane Conductance Regulator/metabolismElectrophysiologyEpithelial Cells/cytology/metabolismFibroblasts/cytology/metabolismHumansInterleukin-8/metabolismMucins/metabolismRespiratory Mucosa/cytologyStem Cells
Identifiers
PMID: 16179582
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Research group Mucoviscidose et jonctions GAP (229)
Projects
Swiss National Science Foundation: 3100-067120.01
Swiss National Science Foundation: 3100A0-100621-1
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(ISO format)
WISZNIEWSKI, Ludovic et al. Long-term cultures of polarized airway epithelial cells from patients with cystic fibrosis. In: American journal of respiratory cell and molecular biology, 2006, vol. 34, n° 1, p. 39-48. doi: 10.1165/rcmb.2005-0161OC https://archive-ouverte.unige.ch/unige:27057

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Deposited on : 2013-03-26

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