Advertisement

Well-Differentiated Human Airway Epithelial Cell Cultures

  • M. Leslie Fulcher
  • Sherif Gabriel
  • Kimberlie A. Burns
  • James R. Yankaskas
  • Scott H. Randell
Part of the Methods in Molecular Medicine™ book series (MIMM, volume 107)

Abstract

The airway epithelium occupies a critical environmental interface, protecting the host from a wide variety of inhaled insults, including chemical and particulate pollutants and pathogens. The coordinated regulation of ion and water transport, mucous secretion, and cilia beating underlies mucociliary clearance. Physical trapping and removal of harmful substances, in combination with baseline or inducible secretion of antimicrobial factors, antioxidants, and protease inhibitors and recruitment of nonspecific inflammatory cells (neutrophils, monocytes), constitutes airway innate host defense.

References

  1. 1.
    Lechner, J. R, Haugen, A., McLendon, I. A., and Pettis, E. W. (1982) Clonal growth of normal adult human bronchial epithelial cells in a serum-free medium. In Vitro 18, 633–642.PubMedCrossRefGoogle Scholar
  2. 2.
    Gruenert, D. C., Finkbeiner, W. E., and Widdicombe, J. H. (1995) Culture and transformation of human airway epithelial cells. Am. J. Physiol. 268, L347–L360.PubMedGoogle Scholar
  3. 3.
    Terzaghi, M., Nettesheim, P., and Williams, M.L. (1978) Repopulation of denuded tracheal grafts with normal, preneoplastic, and neoplastic epithelial cell populations. Cancer Res. 38, 4546–4553.PubMedGoogle Scholar
  4. 4.
    Yankaskas, J. R., Knowles, M. R., Gatzy, J. T., and Boucher, R. C. (1985) Persistence of abnormal chloride ion permeability in cystic fibrosis nasal epithelial cells in heterologous culture. Lancet 1, 954–956.PubMedCrossRefGoogle Scholar
  5. 5.
    Jorissen, M., Van Der Schueren, B., van den Berghe, H., and Cassiman, J. J. (1989) The preservation and regeneration of cilia on human nasal epithelial cells cultured in vitro. Arch. Otorhinolaryngol. 246, 308–314.PubMedCrossRefGoogle Scholar
  6. 6.
    Wu, R., Yankaskas, J., Cheng, E., Knowles, M. R., and Boucher, R. (1985) Growth and differentiation of human nasal epithelial cells in culture. Serum-free, hormone-supplemented medium and proteoglycan synthesis. Am. Rev. Respir. Dis. 132, 311–320.PubMedGoogle Scholar
  7. 7.
    Benali, R., Tournier, J. M., Chevillard, M., et al. (1993) Tubule formation by human surface respiratory epithelial cells cultured in a three-dimensional collagen lattice. Am. J. Physiol. 264, L183–L192.PubMedGoogle Scholar
  8. 8.
    Whitcutt, M. J., Adler, K., and Wu, R. (1988) A biphasic chamber system for maintaining polarity of differentiation of cultured respiratory tract epithelial cells. In Vitro Cell. Dev. Biol. 24, 420–428.PubMedCrossRefGoogle Scholar
  9. 9.
    Matsui, H., Grubb, B. R., Tarran, R., et al. (1998) Evidence for periciliary liquid layer depletion, not abnormal ion composition, in the pathogenesis of cystic fibrosis airways disease. Cell 95, 1005–1015.PubMedCrossRefGoogle Scholar
  10. 10.
    Matsui, H., Randell, S. H., Peretti, S. W., Davis, C. W., and Boucher, R. C. (1998) Coordinated clearance of periciliary liquid and mucus from airway surfaces. J. Clin. Invest. 102, 1125–1131.PubMedCrossRefGoogle Scholar
  11. 11.
    Pickles, R. J., McCarty, D., Matsui, H., Hart, P. J., Randell, S. H., and Boucher, R. C. (1998) Limited entry of adenoviral vectors into well differentiated airway epithelium is responsible for inefficient gene transfer. J. Virol. 72, 6014–6023.PubMedGoogle Scholar
  12. 12.
    Zhang, L., Peeples, M. E., Boucher, R. C., Collins, P. L., and Pickles, R. J. (2002) Respiratory syncytial virus infection of human airway epithelial cells is polarized, specific to ciliated cells, and without obvious cytopathology. J. Virol. 76, 5654–5666.PubMedCrossRefGoogle Scholar
  13. 13.
    Bernacki, S. H., Nelson, A. L., Abdullah, L., et al. (1999) Mucin gene expression during differentiation of human airway epithelia in vitro. Muc4 and muc5b are strongly induced. Am. J. Respir. Cell Mol. Biol. 20, 595–604.PubMedGoogle Scholar
  14. 14.
    Zhang, Y. J., O’Neal, W. K., Randell, S. H., et al. (2002) Identification of dynein heavy chain 7 as an inner arm component of human cilia that is synthesized but not assembled in a case of primary ciliary dyskinesia. J. Biol. Chem. 277, 17,906–17,915.PubMedCrossRefGoogle Scholar
  15. 15.
    Yoon, J. H., Koo, J. S., Norford, D., Guzman, K., Gray, T., and Nettesheim, P. (1999) Lysozyme expression during metaplastic squamous differentiation of retinoic acid-deficient human tracheobronchial epithelial cells. Am. J. Respir. Cell Mol. Biol. 20, 573–581.PubMedGoogle Scholar
  16. 16.
    Gray, T. E., Guzman, K., Davis, C. W., Abdullah, L. H., and Nettesheim, P. (1996) Mucociliary differentiation of serially passaged normal human tracheobronchial epithelial cells. Am. J. Respir. Cell Mol. Biol. 14, 104–112.PubMedGoogle Scholar
  17. 17.
    Lechner, J. F. and LaVeck, M. A. (1985) A serum-free method for culturing normal human bronchial epithelial cells at clonal density. J. Tiss. Cult. Meth. 9, 43–48.CrossRefGoogle Scholar
  18. 18.
    Randell, S. H., Walstad, L., Schwab, U. E., Grubb, B. R., and Yankaskas, J. R. (2001) Isolation and culture of airway epithelial cells from chronically infected human lungs. In Vitro Cell Dev. Biol. Anim. 37, 480–489.PubMedCrossRefGoogle Scholar
  19. 19.
    Bancroft, J. and Stevens, A. (1996) Theory and Practice of Histological Techniques, Battle Press, Columbus, OH, pp. 69–80.Google Scholar
  20. 20.
    Sheehan, D. and Hrapchak, B. (1980) Theory and Practice of Histotechnology, Battle Press, Columbus, OH, Vol. 2, 59–66.Google Scholar
  21. 21.
    Hayat, M. A. (1989) Principles and Techniques of Electron Microscopy 3, 79–92.Google Scholar
  22. 22.
    Goldstein, J. (1984) Scanning Electron Microscopy and X-ray Micro analysis, pp. 495–540.Google Scholar
  23. 23.
    Boat, T. E, Welsh, M. J., and Beaudet, A. L. (1989) Cystic fibrosis, in The Metabolic Basis of Inherited Disease (Scriver, C. R., Beaudet, A. L., Sly, W. S., Valle, D., Stansbury, J. B., Wyngaarden, J. B., and Frederickson, D. S., eds.), 2649–2680.Google Scholar
  24. 24.
    Rommens, J. M., Iannuzzi, M. C., Kerem, B.-T., et al. (1989) Identification of the cystic fibrosis gene: Chromosome walking and jumping. Science 245, 1059–1065.PubMedCrossRefGoogle Scholar
  25. 25.
    Riordan, J. R., Rommens, J. M., Kerem, B.-T., et al. (1989) Identification of the cystic fibrosis gene: Cloning and characterization of complementary DNA. Science 245, 1066–1073.PubMedCrossRefGoogle Scholar
  26. 26.
    Kerem, B., Rommens, J. M., Buchanan, J. A., et al. (1989) Identification of the cystic fibrosis gene: Genetic analysis. Science 245, 1073–1080.PubMedCrossRefGoogle Scholar
  27. 27.
    Anderson, M. P., Rich, D. P., Gregory, R. J., Smith, A. E., and Welsh, M. J. (1991) Generation of cAMP-activated chloride currents by expression of CFTR. Science 251, 679–682.PubMedCrossRefGoogle Scholar
  28. 28.
    Kartner, N., Hanrahan, J. W., Jensen, T. J., et al. (1991) Expression of the cystic fibrosis gene in non-epithelial invertebrate cells produces a regulated anion conductance. Cell 64, 681–691.PubMedCrossRefGoogle Scholar
  29. 29.
    Stutts, M. J., Canessa, C. M., Olsen, J. C., et al. (1995) CFTR as a cAMP-dependent regulator of sodium channels. Science 269, 847–850.PubMedCrossRefGoogle Scholar
  30. 30.
    Gabriel, S. E., Makhlina, M., Martsen, E., Thomas, E. J., Lethem, M. L, and Boucher, R. C. (2000) Permeabilization via the P2X7 purinoceptor reveals the presence of a Ca2+-activated Cl- conductance in the apical membrane of murine tracheal epithelial cells. J. Biol. Chem. 275, 35,028–35,033.PubMedCrossRefGoogle Scholar
  31. 31.
    Tarran, R., Loewen, M. E., Paradiso, A. M., et al. (2002) Regulation of murine airway surface liquid volume by CFTR and Ca2+-activated Cl- conductances. J. Gen. Physiol. 120, 407–418.PubMedCrossRefGoogle Scholar
  32. 32.
    Donaldson, S. H., Hirsh, A., Li, D. C., et al. (2002) Regulation of the epithelial sodium channel by serine proteases in human airways. J. Biol. Chem. 277, 8338–8345.PubMedCrossRefGoogle Scholar

Copyright information

© Humana Press Inc. 2005

Authors and Affiliations

  • M. Leslie Fulcher
    • 1
  • Sherif Gabriel
    • 1
  • Kimberlie A. Burns
    • 1
  • James R. Yankaskas
    • 1
  • Scott H. Randell
    • 1
  1. 1.Cystic Fibrosis/Pulmonary Research and Treatment CenterThe University of North Carolina at Chapel HillUSA

Personalised recommendations