Summary
Tracheal epithelial cells were grown on Nuclepore filters coated with human placental collagen. When grown immersed in medium containing fetal bovine serum, cells displayed an undifferentiated ultrastructure (no cilia and a cell height of ∼ 10 µm). Short-circuit current (Isc) was approximately 1/10 that of the native epithelium. By contrast, when grown in hormonally defined, serum-free medium with an air interface, cells showed Isc equal to or greater than the original tissue, possessed cilia, and had a cell height of ∼ 50 µm. Responses in Isc to mediators were similar to those of the original tissue, but differed from those of dog or human tracheal epithelium. Given the ready availability and low cost of the native tissues, bovine tracheal cultures grown in serum-free medium with an air interface should prove useful in studies of airway epithelial physiology.
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References
Al-Bazzaz, F. J.; Cheng, E. Effect of catecholamines on ion transport in dog tracheal epithelium. J. Appl. Physiol. 47:397–403; 1979.
Boucher, R. C.; Larsen, E. H. Comparison of ion transport by cultured secretory and absorptive canine airway epithelia. Am. J. Physiol. 254:C535-C547; 1988.
Cesarone, C. F.; Bolognesi, C.; Santi, L. Improved microfluorometric DNA determination in biological material using Hoechst 33258. Anal. Biochem. 100:188–197; 1979.
Coleman, D. L.; Tuet, I. K.; Widdicombe, J. H. Electrical properties of dog tracheal epithelial cells grown in monolayer culture. Am. J. Physiol. 246:C355-C359; 1984.
Cotton, C. U.; Lawson, E. E.; Boucher, R. C., et al. Bioelectric properties and ion transport of airways excised from adult and fetal sheep. J. Appl. Physiol. 55:1542–1549; 1983.
Hartmann, T.; Kondo, M.; Mochizuki, H., et al. Calcium-dependent regulation of C1 secretion in tracheal epithelium. Am. J. Physiol. 262:L163-L168; 1992.
Kondo, M.; Finkbeiner, W. E.; Widdicombe, J. H. A simple technique for culture of highly differentiated cells from dog tracheal epithelium. Am. J. Physiol. 216:L106-L117; 1991.
Langridge-Smith, J. E.; Rao, M. C.; Field, M. Chloride and sodium transport across bovine tracheal epithelium: effects of secretagogues and indomethacin. Pfluegers Arch. 402:42–47; 1984.
Schumann, B. L.; Cody, T. E.; Miller, M., et al. Isolation, characterization, and long-term culture of fetal bovine tracheal epithelial cells. In Vitro Cell. Dev. Biol. 24:211–216; 1988.
Smith, P. K.; Krohn, R. I.; Hermanson, G. T., et al. Measurement of protein using bicinchoninic acid. Anal. Biochem. 71:1392–1401; 1985.
Van Scott, M. R.; Lee, N. P.; Yankaskas, J. R., et al. Effect of hormones on growth and function of cultured canine tracheal epithelial cells. Am. J. Physiol. 255:C237-C245; 1988.
Vulliemin, P.; Durand-Arczynska, W.; Durand, J. Electrical properties and electrolyte transport in bovine tracheal epithelium: effects of ion substitutions, transport inhibitors and histamine. Pfluegers Arch. 396:54–59; 1983.
Widdicombe, J. H. Electrical methods for studying ion and fluid transport across airway epithelia. In: Allegra, L.; Braga, P., eds. Methods in bronchial mucology. New York: Raven Press; 1988:335–345.
Widdicombe, J. H. Use of cultured airway epithelial cells in studies of ion transport. Am. J. Physiol. 258:L13-L18; 1990.
Widdicombe, J. H.; Basbaum, C. B.; Highland, E. Ion contents and other properties of isolated cells from dog tracheal epithelium. Am. J. Physiol. 241:C184-C192; 1981.
Widdicombe, J. H.; Coleman, D. L.; Finkbeiner, W. E., et al. Primary cultures of the dog’s tracheal epithelium: fine structure, fluid and electrolyte transport. Cell Tissue Res. 247:94–103; 1987.
Widdicombe, J. H.; Coleman, D. L.; Finkbeiner, W. E., et al. Electrical properties of monolayers cultured from cells of human tracheal mucosa. J. Appl. Physiol. 58:1729–1735; 1985.
Wu, T.; Yankaskas, J. R.; Cheng, E., et al. 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; 1985.
Yamaya, M.; Finkbeiner, W. E.; Chun, S. Y., et al. Differentiated structure and function of cultures from human tracheal epithelium. Am. J. Physiol. 262:L713-L724; 1992.
Zeitlin, P. L.; Loughlin, G. M.; Guggino, W. B. Ion transport in cultured fetal and adult rabbit epithelia. Am. J. Physiol. 254:C691-C698; 1988.
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Kondo, M., Finkbeiner, W.E. & Widdicombe, J.H. Cultures of bovine tracheal epithelium with differentiated ultrastructure and ion transport. In Vitro Cell Dev Biol - Animal 29, 19–24 (1993). https://doi.org/10.1007/BF02634367
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DOI: https://doi.org/10.1007/BF02634367