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Effects of ouabain on the function and structure of a cell line (MDCK) derived from canine kidney

I. Light microscopic observations of monolayer growth

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Summary

MDCK is a transformed cell line derived from normal dog kidney. It exhibits qualities of renal tubular epithelium and of papillary adenocarcinoma. On solid substrates, MDCK forms confluent monolayers that are studded with multicellular blisters and cordlike structures. Ouabain, at a concentration of 4×10−8 m, abolishes blister formation within 24 hr; withdrawal of the drug brings about reappearance of the blisters. Ouabain-sensitive ATPase activity has been demonstrated in these cells by biochemical measurement. These findings suggest that blister formation in MDCK monolayer growth is associated with active transport of solutes accompanied by water and that 4×10−8 m of ouabain interferes with these transport systems. As an additional early effect, the drug decreases the frequency of mitosis.

The exposure of MDCK monolayers to ouabain for several weeks results in profound cytological alterations. A marked eosinophilic stippling of the periphery of chronically exposed cells is seen with the light microscope. After prolonged exposure to ouabain, the cell population of MDCK monolayers becomes clearly heterogeneous, with frequent giant cells and foci of small, hyperchromatic proliferating cells. The small cells may indicate the appearance of ouabain-resistant strains.

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This investigation was part of a dissertation submitted to the University of Pittsburgh in partial fulfillment of the requirements for the degree of Doctor of Philosophy.

This study was supported by NIH-NCI Research Grants CA 13219 and CA 14137, by Contract G-72-3858, and by NIH Fellowship FO 3 DE 426160 from the National Institutes of Health.

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Abaza, N.A., Leighton, J. & Schultz, S.G. Effects of ouabain on the function and structure of a cell line (MDCK) derived from canine kidney. In Vitro 10, 172–183 (1974). https://doi.org/10.1007/BF02615230

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