A Method for the Preparation of Suspensions of Intestinal Mucosal Cells by Means of Calcium Chelators
(A preliminary report)
Acta Veterinaria Scandinavica volume 8, pages 76–82 (1967)
A method is described for the preparation of a suspension of single cells of the intestinal mucosa by the calcium chelators ED TA and EGTA.
Preliminary experiments with oxygen consumption and glucose-utilization indicate a considerable metabolic activity of the cell suspension.
The difference in glycine uptake in presence and absence of sodium and the effect of phlorizin on the glucose metabolism of the cells indicates the existence of a qualitatively normal membrane function.
The application of a similar method for the purpose of obtaining viable in vitro preparations from intestinal tissue of larger animals is suggested.
Bronk, J. B. & D. S. Parsons: Accumulation and incorporation of amino acid in rat intestine in vitro. Nature (Lond.) 1965, 208, 786–787.
Clark, B. & J. W. Porteous: The isolation and properties of epithelial cell “ghosts” from rat small intestine. Biochem. J. 1965, 96, 539–551.
Erichsen, S. & E. Søgnen: Unpublished data 1964.
Foreman, H. & T. T. Trujillo: The metabolism of C14-labeled ethylene-diaminetetraacetic acid in human beings. J. Lab. clin. Med. 1954, 43, 566–571.
Hansen, I. A.: Effects of Ga++ and Mg++ on cell adhesion. J. Sci. indust. Res. 1959, 180, 84–85.
Harrer, D. S., B. K. Stern & R. W. Reilly: Removal and dissociation of epithelial cells from the rodent gastrointestinal tract. Nature (Lond.) 1964, 203, 319–320.
Newey, H., B. J. Parsons & D. H. Smyth: The site of action of phlorrhi-zin in inhibiting intestinal absorption of glucose. J. Physiol. (Lond.) 1959, 148, 83–92.
Riklis, E. & J. H. Quastel: Effects of cations on sugar absorption by isolated surviving guinea pig intestine. Canad. J. Biochem. 1958, 36, 347–362.
Stern, B. K. & R. W. Reilly: Some characteristics of the respiratory metabolism of suspensions of rat intestinal cells. Nature (Lond.) 1965, 205, 563–565.
Stern, B. K. & W. E. Jensen: Active transport of glucose by suspensions of isolated rat intestinal cells. Nature (Lond.) 1966, 209, 789–790.
Stirling, C. E. & W. B. Vinter: Phlorizin-H3 distribution in mucosa from hamster intestine during inhibition of sugar transport. Fed. Proc. 50th Ann. Meet. 1966, 25, Part 1, 640.
Søgnen, E.: Intestinal transport of glucose and sulphanilamide in vitro at low calcium concentrations and in the presence of calcium-binding substances. Acta pharcnacol. (Kbh.) 1965, 22, 61–68.
Tenenhouse, A. & J. H. Quastel: Amino acid accumulation in Ehrlich ascites carcinoma cells. Canad. J. Biochem. 1960, 38, 1311–1326.
Wilson, T. H. & G. Wiseman: The use of sacs of everted small intestine for the study of the transference of substances from the mucosal to the serosal surface. J. Physiol. (Lond.) 1954, 123, 116–125.
Wilson, T. H.: Intestinal Absorption. W. B. Saunders, London 1962, p. 34.
I wish to thank Professor J. H. Quastel for the opportunity to work in his unit and for valuable advice from him and his coworkers. Thanks are also due to Professor C. P. Leblond and Mr. G. Altmann for their preliminary histological examination of the cells, and to Mr. Joav Prives for his interest in the biochemical aspects of the preparation. My stay in Canada has partly been financed by a grant from the Norwegian Research Council for Sciences and Humanities.
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Søgnen, E. A Method for the Preparation of Suspensions of Intestinal Mucosal Cells by Means of Calcium Chelators. Acta Vet Scand 8, 76–82 (1967). https://doi.org/10.1186/BF03547854