Abstract
Our aim was to develop a fibroblast-free monolayer culture of human gastric mucosal cells, using the specimens obtained by routine endoscopic biopsy. Human gastric mucosa obtained from normal volunteers by endoscopic biopsy was dissociated from collagenase and hyaluronidase. Dissociated cells were cultured in supplemented Coon's modified Ham's F-12 medium. Within 24 hr of inoculation, the cells were attached to the culture dishes. This was followed by cellular outgrowth. On phase-contrast microscopy, all cells had epithelial characteristics and fibroblasts were not observed. Ninety percent of cells contained periodic acid Schiff reaction-positive mucous granules after diastase digestion consistent with mucous epithelial cells. Two percent of the cells gave a strong reaction for succinic dehydrogenase activity (parietal cells). Immunohistochemical staining for pepsinogen in cultured cells was negative. On EM, microvilli-like projections, junctional complexes, Golgi apparatus, and mucous granules were apparent in the majority of cells. Mitotic figures were observed by day 3 with Giemsa staining. Autoradiographically, these cells were able to incorporate [3H] TdR into the nuclei. Cells were capable of synthesizing DNA, and this function was inhibited by cycloheximide. Cells could be cultured for up to two weeks without fibroblast contamination. A method of primary monolayer culture of human gastric mucosa obtained by a routine endoscopic biopsy has been successfully developed.
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Deschryver-Kecskemeti K, Greider NH, Sake MK, Rieders E, McGuigan JE: The gastric-producing cells in tissue cultures of the rat pyloric antrum. Lab Invest 37:406–410, 1977
Terano A, Ivey KJ, Stachura J, Sekhon S, Hosojima H, McKenzie WN, Krause WJ, Wyche JH: Cell culture of rat gastric fundic mucosa. Gastroenterology 83:1280–1291, 1982
Miller LR, Jacobson ED, Johnson LR: Effect of pentagastrin on gastric mucosal cells grown in tissue culture. Gastroenterology 64:254–267, 1973
Coon HG, Weiss MC: A quantitative comparison of formation of spontaneous and virus-produced viable hybrids. Proc Natl Acad Sci USA 62:852–859, 1969
Phillips HJ: Dye exclusion tests for cell viability.In Tissue Culture Methods and Applications. PF Krause, MD Patterson (eds). New York, Academic Press, 1973, pp 406–408
Nachlas MM, Tsou KC, Souza ED, Cheng CS, Seligman AM: Cytochemical demonstration of succinic dehydrogenase by the use of a newp-nitrophenyl-substituted diterazole. J Histochem Cytochem 5:420–436, 1957
Samloff IM, Liebman WM: Cellular localization of the group II pepsinogens in human stomach and duodenum by immunofluorescence. Gastroenterology 65:36–42, 1973
Paul J: Cell and Tissue Culture, 5th ed. Edinburgh, Churchill Livingstone, 1975, pp 363–364
Bradford M: A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254, 1976
Chopra DP, Yeh KY: Long term culture of epithelial cells from the normal rat colon. In Vitro 17:441–449, 1981
Ito S: Anatomic structure of the gastric mucosa.In Handbook of Physiology: Alimentary Canal. CF Code (ed). Washington, DC, American Physiology Society, 1967, pp 705–741
Gilbert SF, Migeon BR:d-Valine as a selective agent for normal human and rodent epithelial cells in culture. Cell 5:11–17, 1975
Owens RB: Selective cultivation of mammalian epithelial cells.In Methods of Cell Biology. DM Prescott (ed). New York, Academic Press, 1976, pp 341–355
Quaroni A, Wants J, Trestad RL, Isselbacher KJ: Epithelioid cell culture from rat small intestine. J Cell Biol 80:248–265, 1979
Webber MM: Normal and benign human prostatic epithelium in culture. I. Isolation. In Vitro 15:967–982, 1979
White MT, Hu ASL, Hamamoto ST, Nandi S: In vitro analysis of proliferating epithelial cell populations from the mouse mammary gland: Fibroblast-free growth and serial passage. In Vitro 14:271–281, 1978
Lasfargues EY, Moore DH: A method for the continuous cultivation of mammary epithelium. In Vitro 7:21–25, 1971
Lasfargues EY: Collagenase as a cell dispersion agent in tissue cultures.In Collagenase, 1st Interdisciplinary Symposium on Collagenase. I Mandl (ed). New York, Gordon and Breach, 1972, pp 83–89
Paul J: Cell and Tissue Culture, 5th ed. Edinburgh, Churchill Livingstone, 1975, pp 19–22
Helander HF: Ultrastructure and function of gastric parietal cells in the rat during development. Gastroenterology 56:35–52, 1969
Helander HF: Ultrastructure and function of gastric mucoid and zymogen cells in the rat during development. Gastroenterology 56:53–70, 1969
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This work was supported in part by the Medical Research Service of the Veterans Administration. The paper was presented at the annual meeting of the American Gastroenterological Association, Chicago, May 1982.
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Terano, A., Mach, T., Stachura, J. et al. A monolayer culture of human gastric epithelial cells. Digest Dis Sci 28, 595–603 (1983). https://doi.org/10.1007/BF01299919
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DOI: https://doi.org/10.1007/BF01299919