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. 1971 Nov 1;51(2):452–464. doi: 10.1083/jcb.51.2.452

STUDIES ON EPITHELIAL CELLS ISOLATED FROM GUINEA PIG SMALL INTESTINE

E M Evans 1, J M Wrigglesworth 1, K Burdett 1, W F R Pover 1
PMCID: PMC2108144  PMID: 5000170

Abstract

Sheets of mucosal epithelial cells were released from guinea pig small intestine after incubation with ethylenediaminetetraacetate. Cells in sheets retained their columnar shape for 24 hr at room temperature, and exclusion of nigrosine suggested they had intact plasma membranes. When sheets were disaggregated individual cells had normal morphology for at least 4 hr. During isolation 16% of the total protein and 24% of the total lactic dehydrogenase were lost from the cells, but subsequent enzyme leakage was low. Leakage increased with shaking, incubation at 37°C, or increasing the oxygen tension of the suspending medium, but was minimal when the Na+:K+ ratio in the medium was 8:1 and the osmolarity was high. Losses of particulate enzyme activities were negligible. Respiration was constant for up to 4 hr and was insensitive to calcium, bicarbonate, oxygen tension, and pH. It was inhibited by cyanide and iodoacetate and varied with the Na+:K+ ratio of the extracellular fluid and the structural integrity of the cells. All preparations concentrated potassium and excluded sodium, but lost this ability if ouabain was added or cells were broken. Potassium-42 uptake was also sensitive to temperature, ouabain, and structural integrity. The preparations are being used to study cell metabolism in the intestinal epithelium.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. BROWN M. M., PARSONS D. S. Observations on the changes in the potassium content of rat jejunal mucosa during absorption. Biochim Biophys Acta. 1962 May 7;59:249–251. doi: 10.1016/0006-3002(62)90728-x. [DOI] [PubMed] [Google Scholar]
  2. CHANCE B. THE ENERGY-LINKED REACTION OF CALCIUM WITH MITOCHONDRIA. J Biol Chem. 1965 Jun;240:2729–2748. [PubMed] [Google Scholar]
  3. Clark B., Porteous J. W. The isolation and properties of epithelial-cell "ghosts" from rat small intestine. Biochem J. 1965 Aug;96(2):539–551. doi: 10.1042/bj0960539. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. DALTON A. J., FELIX M. D. Studies on the Golgi substance of the epithelial cells of the epididymis and duodenum of the mouse. Am J Anat. 1953 Mar;92(2):277–305. doi: 10.1002/aja.1000920204. [DOI] [PubMed] [Google Scholar]
  5. Exton J. H. Metabolism of rat-liver cell suspensions. 1. General properties of isolated cells and occurrence of the citric acid cycle. Biochem J. 1964 Sep;92(3):457–467. doi: 10.1042/bj0920457. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. GIANETTO R., DE DUVE C. Tissue fractionation studies. 4. Comparative study of the binding of acid phosphatase, beta-glucuronidase and cathepsin by rat-liver particles. Biochem J. 1955 Mar;59(3):433–438. doi: 10.1042/bj0590433. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. GLOCK G. E., McLEAN P. Further studies on the properties and assay of glucose 6-phosphate dehydrogenase and 6-phosphogluconate dehydrogenase of rat liver. Biochem J. 1953 Oct;55(3):400–408. doi: 10.1042/bj0550400. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Hampton J. C., Rosario B. The attachment of microorganisms to epithelial cells in the distal ileum of the mouse. Lab Invest. 1965 Aug;14(8):1464–1481. [PubMed] [Google Scholar]
  9. Harrison D. D., Webster H. L. The preparation of isolated intestinal crypt cells. Exp Cell Res. 1969 May;55(2):257–260. doi: 10.1016/0014-4827(69)90489-3. [DOI] [PubMed] [Google Scholar]
  10. Huang K. C. Uptake of L-tyrosine and 3-0-methylglucose by isolated intestinal epithelial cells. Life Sci. 1965 Jun;4(12):1201–1206. doi: 10.1016/0024-3205(65)90333-4. [DOI] [PubMed] [Google Scholar]
  11. Hübscher G., West G. R., Brindley D. N. Studies on the fractionation of mucosal homogenates from the small intestine. Biochem J. 1965 Dec;97(3):629–642. doi: 10.1042/bj0970629. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Iemhoff W. G., van den Berg J. W., de Pijper A. M., Hülsmann W. C. Metabolic aspects of isolated cells from rat small intestinal epithelium. Biochim Biophys Acta. 1970 Aug 14;215(2):229–241. doi: 10.1016/0304-4165(70)90020-6. [DOI] [PubMed] [Google Scholar]
  13. KALTENBACH J. P., KALTENBACH M. H., LYONS W. B. Nigrosin as a dye for differentiating live and dead ascites cells. Exp Cell Res. 1958 Aug;15(1):112–117. doi: 10.1016/0014-4827(58)90067-3. [DOI] [PubMed] [Google Scholar]
  14. Kimmich G. A. Active sugar accumulation by isolated intestinal epithelial cells. A new model for sodium-dependent metabolite transport. Biochemistry. 1970 Sep 15;9(19):3669–3677. doi: 10.1021/bi00821a004. [DOI] [PubMed] [Google Scholar]
  15. Kimmich G. A. Preparation and properties of mucosl epithelial cells isolated frmsmall intestine of the chicken. Biochemistry. 1970 Sep 15;9(19):3659–3668. doi: 10.1021/bi00821a003. [DOI] [PubMed] [Google Scholar]
  16. PENNINGTON R. J. Biochemistry of dystrophic muscle. Mitochondrial succinate-tetrazolium reductase and adenosine triphosphatase. Biochem J. 1961 Sep;80:649–654. doi: 10.1042/bj0800649. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Perris A. D. Isolation of the epithelial cells of the rat small intestine. Can J Biochem. 1966 Jun;44(6):687–693. doi: 10.1139/o66-086. [DOI] [PubMed] [Google Scholar]
  18. REIMANN H. A. MICROBIC PHAGOCYTOSIS BY ENTERIC EPITHELIAL CELLS. JAMA. 1965 Jun 28;192:1130–1132. doi: 10.1001/jama.1965.03080260018005. [DOI] [PubMed] [Google Scholar]
  19. RIKLIS E., QUASTEL J. H. Effects of cations on sugar absorption by isolated surviving guinea pig intestine. Can J Biochem Physiol. 1958 Mar;36(3):347–362. [PubMed] [Google Scholar]
  20. Rochman H., Clark P. B., Lathe G. H., Parsons F. M. The effect of temperature and anoxia of rat-kidney slices on their subsequent respiration. Biochem J. 1967 Jan;102(1):44–47. doi: 10.1042/bj1020044. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. SCHAEDLER R. W., DUBS R., COSTELLO R. ASSOCIATION OF GERMFREE MICE WITH BACTERIA ISOLATED FROM NORMAL MICE. J Exp Med. 1965 Jul 1;122:77–82. doi: 10.1084/jem.122.1.77. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. SCHREINER G. E. Determination of inulin by means of resorcinol. Proc Soc Exp Biol Med. 1950 May;74(1):117–120. doi: 10.3181/00379727-74-17827. [DOI] [PubMed] [Google Scholar]
  23. SHERRATT H. S., HUBSCHER G. Properties of mitochondrial preparations from the small-intestinal mucosa of the guinea-pig. Biochim Biophys Acta. 1963 Feb 5;69:403–405. doi: 10.1016/0006-3002(63)91274-5. [DOI] [PubMed] [Google Scholar]
  24. SKOU J. C. ENZYMATIC BASIS FOR ACTIVE TRANSPORT OF NA+ AND K+ ACROSS CELL MEMBRANE. Physiol Rev. 1965 Jul;45:596–617. doi: 10.1152/physrev.1965.45.3.596. [DOI] [PubMed] [Google Scholar]
  25. Schultz S. G., Fuisz R. E., Curran P. F. Amino acid and sugar transport in rabbit ileum. J Gen Physiol. 1966 May;49(5):849–866. doi: 10.1085/jgp.49.5.849. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Sjöstrand F. S. A simple and rapid method to prepare dispersions of columnar epithelial cells from the rat intestine. J Ultrastruct Res. 1968 Mar;22(5):424–442. doi: 10.1016/s0022-5320(68)90032-4. [DOI] [PubMed] [Google Scholar]
  27. Stern B. K., Jensen W. E. Active transport of glucose by suspensions of isolated rat intestinal epithelial cells. Nature. 1966 Feb 19;209(5025):789–790. doi: 10.1038/209789a0. [DOI] [PubMed] [Google Scholar]

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