Skip to main content
Log in

Isolation and culturing of highly polarized primary epithelial cells from normal human stomach (antrum) as spheroid-like vesicles

  • Published:
Methods in Cell Science

Abstract

A novel procedure is described for the three-dimensional (3-D) in vitro culture and for maintaining of nontransformed gastric epithelial cells from the human antrum mucosa (HAEC). Biopsies obtained from the antrum were cut into small pieces and the tissue fragments were incubated in culture medium containing the appropriate antibiotics. The suspended mucosal fragments generated small, spheroid-like vesicles consisting of predominantly highly prismatic, mucus-producing cells which mimic the in vivo counterparts structurally and functionally. Electron microscopic investigations revealed a number of ultrastructural and morphological features similar to those of normal gastric cells in vivo such as apical microvilli associated with a glycocalyx, tight junctions, desmosomes, membraneous infoldings, mucous droplets, and an irregular basal lamina. In comparison to the two-dimensional (2-D) gastric cell cultures grown on plane supports, the vesicles maintain an intact epithelial organization of individual cells. The prismatic phenotype, the histophysiology as well as the cytoarchitecture of the non-transformed 3-D cultured gastric epithelial cells are comparable to those of the native tissue and therefore represent a suitable model for defined pathogen-host cell interactions.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  1. Prewett EY, Bickley J, Owen RJ, Pounder RE (1992). DNA patterns of Helicobacter pylori isolated from gastric antrum, body, and duodenum. Gastroenterology 102: 829–833.

    PubMed  Google Scholar 

  2. Bayerdörffer E, Lehn N, Hatz R, Mannes GA, Oertel H, Sauerbruch T, Stolte M (1992). Difference in expression of Helicobacter pylori gastritis in antrum and body. Gastroenterology 102: 1575–1582.

    PubMed  Google Scholar 

  3. Haas R (1995). Mikrobiologie, Pathogenität und molekulare Mechanismen der Virulenz von Helicobacter pylori. Biospektrum 3: 35–40.

    Google Scholar 

  4. Jankowski J, Henderson K, Viaene A, Baert J, Long LQ (1995). Morphological analysis of gastroesophageal diseases by molecular cell techniques. Micros Res Tech 31: 184–192.

    Google Scholar 

  5. Terano A, Mach T, Stachura J, Sekhon S, Tarnawski A, Ivey KJ (1983). A monolayer culture of human gastric epithelial cells. Digest Dis Sci 28: 595–603. 0

    PubMed  Google Scholar 

  6. Moyer MP (1983). Culture of human gastrointestinal epithelial cells. Proc Soc Exp Biol Med 174: 12–15. 0

    PubMed  Google Scholar 

  7. Boyd KL, Forte JG (1994). Primary culture of isolated gastric glands as a model for gastric cell function. FASEB J 8: abstract, No. 3280.

    Google Scholar 

  8. Streuli CH, Bailey N, Bissell MJ (1991). Control of mammary epithelial differentiation: the separate roles of cell-substratum and cell-cell interaction. J Cell Biol 115: 1383–1395.

    Article  PubMed  Google Scholar 

  9. Nelson WJ, Veshnock PJ (1987a). Ankyrin binding to (Na++K+)ATPase and implications for the organization of membrane domains in polarized cells. Nature 328: 533–536.

    Article  PubMed  Google Scholar 

  10. Nelson WJ, Veshnock PJ (1987b). Modulation of Fodrin (membrane skeleton) stability by cell-cell contact in Madin-Darby Canine Kidney epithelial cells. J Cell Biol 104: 1527–1537.

    PubMed  Google Scholar 

  11. Rodriguez-Boulan E, Salas PJI (1989). External and internal signals for epithelial cell surface polarization. Annu Rev Physiol 51: 741–754.

    PubMed  Google Scholar 

  12. Rodriguez-Boulan E, Nelson WJ (1989). Morphogenesis of the polarized epithelial cell phenotype. Science 245: 718–725.

    PubMed  Google Scholar 

  13. Boxberger H-J, Sessler MJ, Maetzel B, Meyer TF (1993). Highly polarized primary epithelial cells from human nasopharynx grown as spheroid-like vesicles. Eur J Cell Biol 62: 140–151.

    PubMed  Google Scholar 

  14. Boxberger H-J, Sessler MJ, Maetzel B, Mosleh IM, Becker H-D, Meyer TF (1994). Highly polarized primary urothelial cells from human ureter grown as spheroid-like vesicles. Epith Cell Biol 3: 85–95.

    Google Scholar 

  15. Boxberger H-J, Sessler MJ, Grund K-E, Meyer TF (1995). Three-dimensional (3-D) cell culture of epithelial cells from the human colonic mucosa. J Exp Clin Cancer Res 14 (Suppl. 1): 22–23.

    Google Scholar 

  16. Buset M, Lipkin M, Winawer S, Swaroop S, Friedman E (1986). Inhibition of human colonic epithelial cell proliferation in vivo and in vitro by calcium. Cancer Research 46: 5426–5430.

    PubMed  Google Scholar 

  17. Kondo Y, Rose I, Young GP, Whitehead RH (1984). Growth and differentiation of fetal rat small intestinal epithelium in tissue culture. Exp Cell Res 153: 121–134.

    PubMed  Google Scholar 

  18. Herzog V, Miller F (1981). Structural and functional polarity of inside-out follicles prepared from pig thyroid gland. Eur J Cell Biol 24: 74–84.

    PubMed  Google Scholar 

  19. König G (1991). Polychloroprene glue as an adhesive for mounting specimens for SEM. J Electron Microsc Tech 17: 467–468.

    PubMed  Google Scholar 

  20. Gibson-D'Ambrosio RE, Samuel M, D'Ambrosio SM (1986). A method for isolating large numbers of viable disaggregated cells from various human tissues for cell culture establishment. In Vitro Cell Dev Biol 22: 529–534.

    PubMed  Google Scholar 

  21. Raul F, Kedinger M, Simon P, Grenier J, Haffen K (1978). Behaviour of isolated rat intestinal cells maintained in suspension or monolayer cultures. Biol Cell 33: 163–168.

    Google Scholar 

  22. Tsuchiya W, Okada Y (1982). Membrane potential changes associated with differentiation of enterocytes in the rat intestinal villi in culture. Dev Biol 94: 284–290.

    PubMed  Google Scholar 

  23. Quaroni A (1985). Development of fetal rat intestine in organ and monolayer culture. J Cell Biol 100: 1611–1622.

    PubMed  Google Scholar 

  24. Minuth WW, Stöckl G, Kloth S, Dermietzel R (1992). Construction of an apparatus for perfusion cell cultures which enables in vitro experiments under organotypic conditions. Eur J Cell Biol 57: 132–137.

    PubMed  Google Scholar 

  25. Owens RB (1976). Selective cultivation of mammalian epithelial cells. In: Prescott DM (ed), Methods of cell biology. New York: Academic Press, pp 341–355.

    Google Scholar 

  26. Chlapowski FJ, Haynes L (1979). The growth and differentiation of transitional epithelium in vitro. J Cell Biol 83: 605–614.

    PubMed  Google Scholar 

  27. Reznikoff CA, Johnson MD, Norback DH, Bryan GT (1983). Growth and characterization of normal human urothelium in vitro. In Vitro 19: 326–343.

    PubMed  Google Scholar 

  28. Walling JM, Blackmore M, Hickman JA, Townsend KMS (1991). Role of the extracellular matrix on the growth and differentiated phenotype of murine colonic adenocarcinoma cells in vitro. Int J Cancer 47: 776–783.

    PubMed  Google Scholar 

  29. Hohn H-P, Denker H-W (1994). The role of cell shape for differentiation of choriocarcinoma cells on extra-cellular matrix. Exp Cell Res 215: 40–50.

    PubMed  Google Scholar 

  30. Nelson WJ, Veshnock PJ (1986). Dynamics of membrane-skeleton (Fodrin) organization during development of polarity in Madin-Darby Canine Kidney epithelial cells. J Cell Biol 103: 1751–1765.

    PubMed  Google Scholar 

  31. McNeil PL (1993). Cellular and molecular adaptions to injurious mechanical stress. Trends Cell Biol 3: 302–307.

    PubMed  Google Scholar 

  32. Karam SH, Leblond CP (1992). Identifying and counting epithelial cell types in the ‘corpus’ of the mouse stomach. Anat Rec 232: 231–246.

    PubMed  Google Scholar 

  33. Mueller-Klieser W (1987). Multicellular spheroids. A review on cellular aggregates in cancer research. J Cancer Res Clin Oncol 113: 101–122.

    PubMed  Google Scholar 

  34. Sutherland RM (1988). Cell and environment interactions in tumor microregions: the multicell spheroid model. Science 240: 177–184.

    PubMed  Google Scholar 

  35. Knüchel R, Sutherland RM (1990). Recent developments in research with human tumor spheroids. The Cancer Journal 3: 234–243.

    Google Scholar 

  36. Négrel R, Rampal P, Nano J-L, Cavenel C, Ailhaud G (1983). Establishment and characterization of an epithelial intestinal cell line from rat fetus. Exp Cell Res 143: 427–437.

    PubMed  Google Scholar 

  37. Jackson CD, Newport G (1981). A simple method for in vitro studies with rodent urinary bladders. In Vitro 17: 107–113.

    PubMed  Google Scholar 

  38. Chevillard M, Hinnrasky J, Pierrot D, Zahm J-M, Klossek J.M, Puchelle E (1993). Differentiation of human surface upper airway epithelial cells in primary culture on a floating collagen gel. Epith Cell Biol 2: 17–25.

    Google Scholar 

  39. Yamanari H, Suganuma T, Iwamura T, Kitamura N, Taniguchi S, Setoguchi T (1994). Extracellular matrix components regulating glandular differentiation and the formation of basal lamina of a human pancreatic cancer cell line in vitro. Exp Cell Res 211: 175–182.

    PubMed  Google Scholar 

  40. Boxberger H-J, Meyer TF (1994). A new method for the 3-D in vitro growth of human RT112 bladder carcinoma cells using the alginate culture technique. Biol Cell 82: 109–119.

    PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Boxberger, HJ., Sessler, M.J., Grausam, M.C. et al. Isolation and culturing of highly polarized primary epithelial cells from normal human stomach (antrum) as spheroid-like vesicles. Methods Cell Sci 19, 169–178 (1997). https://doi.org/10.1023/A:1009751913391

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1023/A:1009751913391

Navigation