Journal of tissue culture methods

, Volume 9, Issue 2, pp 95–105 | Cite as

Isolation, cultivation, and characterization of normal human esophageal epithelial cells

  • Susan P. Banks-Schlegel


Normal human esophageal epithelial cells can now be grown and serially propagated in cell culture either using serum-containing medium and lethally irradiated 3T3 fibroblasts to support epithelial cell growth or using hormone-supplemented, serum-free medium without a 3T3 “feeder” layer. The cells continue to express many differentiated functions characteristic of the native epithelium. When grown in serum-containing medium, the esophageal epithelial cell colonies are stratified and consist of a basal layer of dividing cells giving rise to the more differentiated cells in the upper cell layers. The cells contain keratin proteins and desmosomes between adjacent cells. The cells express other markers of terminal squamous differentiation, namely, involucrin and cross-linked envelopes. When grown in serum-free, hormone-supplemented medium, the colonies remain as a monolayer rather than stratifying because of the low levels of calcium required for optimal growth. Increased levels of calcium or the presence of serum (2.5% or greater) lead to stratification. The cells grown under serum-free conditions continue to express squamous differentiated properties as keratins, desmosomes (decreased numbers in low-calcium medium), involucrin, and cross-linked envelopes.

Key words

human esophagus epithelium cell culture differentiation keratin 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

V. References

  1. 1.
    Banks-Schlegel, S. P.; Green, H. Studies on the development of the definitive cell type of embryonic epidermis using the cross-linked envelope as differentiation marker. Dev. Biol. 74:275–285; 1980.Google Scholar
  2. 2.
    Banks-Schlegel, S. P.; Green, H. Formation of epidermis by serially cultivated human epidermal cells transplanted as an epithelium to athymic mice. Transplantation 29:308–313; 1980.Google Scholar
  3. 3.
    Banks-Schlegel, S. P.; Green, H. Involucrin synthesis and tissue assembly by keratinocytes in natural and cultured human epithelia. J. Cell Biol. 90:732–737; 1981.Google Scholar
  4. 4.
    Banks-Schlegel, S. P.; Harris, C. C. Tissue-specific expression of keratin proteins in human esophageal and epidermal epithelium and their cultured keratinocytes. Exp. Cell Res. 146:271–280; 1983.Google Scholar
  5. 5.
    Banks-Schlegel, S. P.; Howley, P. M. Differentiation of human epidermal cells transformed by SV-40. J. Cell Biol. 96:330–337; 1983.Google Scholar
  6. 6.
    Banks-Schlegel, S. P.; Gazdar, A. F.; Harris, C. C. Intermediate filament and cross-linked envelope expression in human lung tumor cell lines. Cancer Res. 45:1187–1197; 1985.Google Scholar
  7. 7.
    Banks-Schlegel, S. P.; Vocci, M. J.; Combs, J. et al. Normal human esophageal epithelium in cell culture. In: Webber, M.; Sekeley, L., eds. In vitro models for cancer research. Boca Raton: CRC Press; 1985:9–38.Google Scholar
  8. 8.
    Barnes, D.; Sato, G. Methods for growth of cultured cells in serum-free medium. Anal. Biochem. 102:255–270; 1980.Google Scholar
  9. 9.
    Gallico, G. G. III; O'Conner, N. E.; Compton, C. C. et al. Permanent coverage of large burn wounds with autologous cultured human epithelium. N. Engl. J. Med. 311:448–451; 1984.Google Scholar
  10. 10.
    Hennings, H.; Michael, D.; Cheng, C.; et al. Calcium regulation of growth and differentiation of mouse epidermal cells in culture. Cell 19:245–254; 1980.Google Scholar
  11. 11.
    Laemmli, U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685; 1970.Google Scholar
  12. 12.
    Lechner, J. F.; LaVeck, M. A. A serum-free method for culturing normal human bronchial cells at clonal density. J. Tissue Cult. Methods. 9:43–48; 1985.Google Scholar
  13. 13.
    O'Connor, N. E.; Mulliken, J. B.; Banks-Schlegel, S. P. et al. Grafting of burns with cultured epithelium prepared from autologous epidermal cells. Lancet 1:75–78; 1981.Google Scholar
  14. 14.
    Peehl, D. M.; Ham, R. G. Clonal growth of human keratinocytes with small amounts of dialyzed serum. In vitro 16:526–540; 1980.Google Scholar
  15. 15.
    Rheinwald, J. G. Serial cultivation of normal human epidermal keratinocytes. Methods Cell Biol. 21:229–254; 1980.Google Scholar
  16. 16.
    Rice, R. H.; Green, H. Presence in human epidermal cells of a soluble protein precursor of the cross-linked envelope: activation of the cross-linking by calcium ions. Cell 18:681–694; 1979.Google Scholar
  17. 17.
    Rheinwald, J. G.; Green, H. Serial cultivation of strains of human epidermal keratinocytes: the formation of keratinizing colonies from single cells. Cell 6:331–334; 1975.Google Scholar
  18. 18.
    Schlegel, R.; Banks-Schlegel, S. P.; Pinkus, G. S. Immunohistochemical localization of keratin in normal human tissues. Lab. Invest. 42:91–96; 1980.Google Scholar
  19. 19.
    Tsao, M. C.; Walthall, B. J.; Ham, R. G. Clonal growth of normal human epidermal keratinocytes in a defined medium. J. Cell Physiol. 100:219–229; 1982.Google Scholar
  20. 20.
    Watt, F. M. Selective migration of terminally differentiating cells from the basal layer of cultured human epidermis. J. Cell Biol. 98:16–21; 1984.Google Scholar
  21. 21.
    Watt, F. M.; Green, H. Involucrin synthesis is correlated with cell size in human epidermal cultures. J. Cell Biol. 90:738–742; 1981.Google Scholar
  22. 22.
    Watt, F. M.; Green, H. Stratification and terminal differentiation of cultured epidermal cells. Nature 295:434–436; 1982.Google Scholar
  23. 23.
    Watt, F. M.; Mattey, D. L.; Garrod, D. R. Calcium-induced reorganization of desmosomal components in cultured human keratinocytes. J. Cell Biol. 99:2211–2215; 1984.Google Scholar
  24. 24.
    Weinstein, R. Serum-free culture of normal mammalian cells. Biotechnology 1:61–64; 1983.Google Scholar

Copyright information

© Tissue Culture Association, Inc. 1985

Authors and Affiliations

  • Susan P. Banks-Schlegel
    • 1
  1. 1.Laboratory of Human CarcinogenesisNational Cancer Institute, National Institutes of HealthBethesdaUSA

Personalised recommendations