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.
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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.
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.
Haas R (1995). Mikrobiologie, Pathogenität und molekulare Mechanismen der Virulenz von Helicobacter pylori. Biospektrum 3: 35–40.
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.
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
Moyer MP (1983). Culture of human gastrointestinal epithelial cells. Proc Soc Exp Biol Med 174: 12–15. 0
Boyd KL, Forte JG (1994). Primary culture of isolated gastric glands as a model for gastric cell function. FASEB J 8: abstract, No. 3280.
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.
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.
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.
Rodriguez-Boulan E, Salas PJI (1989). External and internal signals for epithelial cell surface polarization. Annu Rev Physiol 51: 741–754.
Rodriguez-Boulan E, Nelson WJ (1989). Morphogenesis of the polarized epithelial cell phenotype. Science 245: 718–725.
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.
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.
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.
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.
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.
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.
König G (1991). Polychloroprene glue as an adhesive for mounting specimens for SEM. J Electron Microsc Tech 17: 467–468.
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.
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.
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.
Quaroni A (1985). Development of fetal rat intestine in organ and monolayer culture. J Cell Biol 100: 1611–1622.
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.
Owens RB (1976). Selective cultivation of mammalian epithelial cells. In: Prescott DM (ed), Methods of cell biology. New York: Academic Press, pp 341–355.
Chlapowski FJ, Haynes L (1979). The growth and differentiation of transitional epithelium in vitro. J Cell Biol 83: 605–614.
Reznikoff CA, Johnson MD, Norback DH, Bryan GT (1983). Growth and characterization of normal human urothelium in vitro. In Vitro 19: 326–343.
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.
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.
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.
McNeil PL (1993). Cellular and molecular adaptions to injurious mechanical stress. Trends Cell Biol 3: 302–307.
Karam SH, Leblond CP (1992). Identifying and counting epithelial cell types in the ‘corpus’ of the mouse stomach. Anat Rec 232: 231–246.
Mueller-Klieser W (1987). Multicellular spheroids. A review on cellular aggregates in cancer research. J Cancer Res Clin Oncol 113: 101–122.
Sutherland RM (1988). Cell and environment interactions in tumor microregions: the multicell spheroid model. Science 240: 177–184.
Knüchel R, Sutherland RM (1990). Recent developments in research with human tumor spheroids. The Cancer Journal 3: 234–243.
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.
Jackson CD, Newport G (1981). A simple method for in vitro studies with rodent urinary bladders. In Vitro 17: 107–113.
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.
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.
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.
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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
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DOI: https://doi.org/10.1023/A:1009751913391