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Isolation, characterization, and long-term culture of fetal bovine tracheal epithelial cells

In Vitro Cellular & Developmental Biology volume 24, pages 211–216 (1988)Cite this article

Summary

Epithelial cells were isolated from fetal bovine trachea by exposing and stripping the mucosal epithelium from the adjacent connective tissue. The tissue was minced and enzymically dissociated in Ca-Mg-free medium containing dispase and dithiothreitol. The stripping procedure and selective trypsinization produced epithelial cell cultures free of fibroblasts. Seeded on plastic, the plating efficiency was 21.5% with a doubling time of 24 h. Dome formation, evidence of occluding junctions and active ion transport characteristic of epithelial cells, was common. Growth of the cells on glass, collagen, and Engelbreth-Holm-Swarm (EHS) substrate demonstrated a striking difference in morphology. Cells grown on EHS presented a more distinctly three-dimensional growth pattern and many more microvilli when compared to cells grown on glass or collagen. The cells retained their epithelioid characteristics through more than 30 passages as shown by the presence of distinct apical and basolateral membranes, tight junctions, and positive keratin staining.

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References

  1. Carney, D. N.; Bunn, P. A.; Gazdar, A. F., et al. Selective growth in serum-free hormone-supplemented medium of tumor cells obtained by biopsy from patients with small cell carcinoma of lung. Proc. Natl. Acad. Sci. USA 78: 3185–3189; 1981.

    PubMed  Article  CAS  Google Scholar 

  2. Coleman, D. L.; Tuet, I. K., Widdicombe, J. H. Electrical properties of dog tracheal epithelial cells grown in monolayer culture. Am. J. Physiol. 246: C355–359; 1984.

    PubMed  CAS  Google Scholar 

  3. Franke, W. W.; Schmid, E.; Osborn, M., et al. Different intermediate-sized filaments distinguished by immunofluorescence microscopy. Proc. Natl. Acad. Sci. USA 75: 5034–5038; 1978.

    PubMed  Article  CAS  Google Scholar 

  4. Freshney, R. I. Culture of animal cells. New York: Alan R. Liss Inc.; 1983.

    Google Scholar 

  5. Gray, T. E.; Thomassen, D. G.; Mass M. J., et al. Quantitation of cell proliferation, colony formation, and carcinogen-induced cytotoxicity of rat tracheal epithelial cells growth in culture on 3T3 feeder layers. In Vitro 19: 559–570; 1983.

    PubMed  Article  CAS  Google Scholar 

  6. Handler, J. S. Use of cultured epithelial to study transport and its regulation. J. Exp. Biol. 106: 55–69; 1983.

    PubMed  CAS  Google Scholar 

  7. Harris, C. O. Human tissues and cells in carcinogenesis research. Cancer Res. 47: 1–10; 1987.

    PubMed  CAS  Google Scholar 

  8. Hay, E. D. Cell biology of extracellular matrix. New York: Plenum Press; 1981.

    Google Scholar 

  9. Lechner, J. F.; Haugen, A.; McClendon, I. A., et al. Clonal growth of normal human bronchial epithelial cells in a serumfree medium. In Vitro 18: 633–642; 1982.

    PubMed  CAS  Google Scholar 

  10. Lee, T. C.; Wu, R.; Brody, A. R., et al. Growth and differentiation of hamster tracheal epithelial cells in culture. Exp. Lung Res. 6: 27–45; 1984.

    PubMed  CAS  Google Scholar 

  11. Leikauf, G. D.; Driscoll, K.; Weideman, P., et al. Ozone-induced augmentation of eicosanoid metabolism in cultured epithelial cells derived from bovine trachea. Fed. Proc. 45: 743; 1986.

    Google Scholar 

  12. Mass, M. J.; Nettesheim, P.; Gray, T. E., et al. The effects of 12-O-tetradecanoylphorbol-13 acetate and other tumor promotors on colony formation of rat tracheal epithelial cells in culture. Carcinogenesis. 5: 1597–1601; 1984.

    PubMed  Article  CAS  Google Scholar 

  13. Milo, G. E.; Ackerman, G. A.; Noyes, I. Growth and ultrastructual characterization of proliferating human keratinocytes in vitro without added extrinsic factors. In Vitro 16: 20–30; 1980.

    PubMed  Article  CAS  Google Scholar 

  14. Misfedt, D. S.; Hamamoto, S. T.; Pitelka, D. R. Transepithelial transport in cell culture. Proc. Natl. Acad. Sci. USA 73: 1212–1216; 1976.

    Article  Google Scholar 

  15. Owens, R. B.; Smith, H. S.; Hackett, A. J. Epithelial cell culture from normal glandular tissue of mice. Mouse epithelial cultures enriched by selective trypsinization. JNCI 53: 261–269; 1974.

    PubMed  CAS  Google Scholar 

  16. Polinger, I. S. Separation of cell types in embryonic heart cell cultures. Exp. Cell Res. 63: 78–82; 1970.

    PubMed  Article  CAS  Google Scholar 

  17. Sun, T. T.; Green, H. Immunofluorescent staining of keratin fibers in cultured cells. Cell 14: 469–476; 1978.

    PubMed  Article  CAS  Google Scholar 

  18. Taub, M.; Sato, G. H. Growth of functional primary cultures of kidney epithelial cells in defined medium. J. Cell Physiol. 105: 369–378; 1980.

    PubMed  Article  CAS  Google Scholar 

  19. Taub, M., ed. Tissue culture of epithelial cells. New York: Plenum Press; 1985.

    Google Scholar 

  20. Thelstad, R. L. The role of extracellular matrix in development. New York: Alan R. Liss Inc.; 1984.

    Google Scholar 

  21. Welsh, M. J. Ion transport by primary cultures of canine tracheal epithelium: methodology, morphology, and electrophysiology. J. Membr. Biol. 88: 149–163; 1985.

    PubMed  Article  CAS  Google Scholar 

  22. 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.

    PubMed  CAS  Google Scholar 

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Affiliations

  1. Department of Environmental Health, University of Cincinnati Medical Center, Mail Location 182, 45267, Cincinnati, OH

    Brenda L. Schumann, Terence E. Cody, Marian L. Miller & George D. Leikauf

  2. Department of Physiology and Biophysics, University of Cincinnati Medical Center, 45267, Cincinnati, Ohio

    George D. Leikauf

Authors
  1. Brenda L. Schumann
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  2. Terence E. Cody
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  3. Marian L. Miller
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  4. George D. Leikauf
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Additional information

This study was supported in part by grants BRSG S07 RR05408-25, Biomedical Research Support Grant Program, Division of Research Resources, by ES 00159, Center Grant, National Institutes of Environmental Health Sciences, by R23-HL37621, New Investigator Award, National Heart, Lung and Blood Institutes, National Institutes of Health, and by the Health Effects Institute, an organization jointly funded by the U.S. Environmental Protection Agency (Assistance Agreement X-8120059) and automotive manufactures. The contents do not necessarily reflect the views of policies of HEI, EPA, or automotive manufacturers.

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Schumann, B.L., Cody, T.E., Miller, M.L. et al. Isolation, characterization, and long-term culture of fetal bovine tracheal epithelial cells. In Vitro Cell Dev Biol 24, 211–216 (1988). https://doi.org/10.1007/BF02623549

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  • DOI: https://doi.org/10.1007/BF02623549

Key words

  • epithelial
  • fetal bovine
  • trachea
  • keratin filaments