Summary
Protein content per cell and cell volume of the canine kidney epithelial cell line, MDCK, and one of its chemically induced tumorigenic transformants (T1) were examined during growth in serum-containing and serum-free, hormonally defined media. Both protein content per cell and cell volume (measured with 3-0-methyl-d-glucose) decreased when cell density increased. Significant inverse linear correlations were found between protein content (and cell volume) and the log of the cell number. Equations relating these variables were derived. No difference was detected between cells grown in serum-containing and in serum-free media. Results of22Na+ uptake experiments differed markedly when expressed as a function of cell number or as a function of protein content or cell volume. However, protein content and cell volume bore a constant relationship to one another, and expression of results as a function of protein content or cell volume yielded qualitatively similar results. These findings show that in MDCK cells, as in other cell lines, protein content and cell volume decrease markedly during growth and that these variation must be taken into account when expressing and interpreting the results of transport measurements.
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Gaush, C. R.; Hard, W. L.; Smith, T. F. Characterization of an established line of canine kidney cells (MDCK). Proc. Soc. Exp. Biol. Med. 122: 931–935; 1966.
Rindler, M. J.; Chuman, L. M.; Shaffer, L.; Saier, M. H., Jr. Retention of differentiated properties in an established dog kidney epithelial cell line (MDCK). J. Cell Biol. 81: 635–648; 1979.
Cereijido, M.; Robbins, E. S.; Dolan, W. J.; Rotunno, C. A.; Sabatini, D. D. Polarized monolayers formed by epithelial cells on permeate and translucent support. J. Cell Biol. 77: 853–880; 1978.
Handler, J. S.; Perkins, F. M.; Johnson, J. P.; Studies of renal cell function using cell culture techniques Am. J. Physiol. 238: F1-F9; 1980.
Misfeldt, D. S.; Hamamoto, S. T.; Pitelka, D. R. Transepithelial transport in cell culture (renal epithelium). Proc. Natl. Acad. Sci. USA 73: 1212–1216; 1976.
Rindler, M. J.; Taub, M.; Saier, M. H., Jr. Uptake of22Na by cultured dog kidney cells. J. Biol. Chem. 254: 11431–11439; 1979.
Taub, M.; Saier, M. H., Jr. Regulation of22Na transport by calcium in an established kidney epithelial cell line. J. Biol. Chem. 254: 11440–11444; 1979.
Taub, M.; Chuman, L.; Saier, M. H., Jr.; Sato, G. Growth of madin-Darby canine kidney epithelial cell (MDCK) line in hormone-supplemented, serum-free medium. Proc. Natl. Acad. Sci. USA 76: 3338–3342; 1979.
Foster, D. O.; Pardee, A. B. Transport of amino acids by confluent and nonconfluent 3T3 and polyoma virus-transformed 3T3 cells growing on glass cover slips. J. Biol. Chem. 244: 2675–2681; 1969.
Kimball, R. F.; Perdue, S. W.; Chu, E. H. Y.; Ortiz, J. R. Microphotometric and autoradiographic studies on the cell cycle and cell size during growth and decline of Chinese hamster cell cultures. Exp. Cell Med. 66: 17–32; 1971.
Kimelberg, H. K.; Mayhew, E. Cell growth and ouabain-sensitive86Rb+ uptake and (Na++K+-ATPase activity in 3T3 and SV40 transformed 3T3 fibroblast. Biochim. Biophys. Acta 45: 865–875; 1976.
Klevecz, R. R.; Ruddle, F. H. Cyclic changes in enzyme activity in synchronized mammalian cell cultures. Science 159: 634–636; 1968.
Mauro, F.; Grasso, A.; Tollmach, L. J. Variations in sulfhydryl, disulfide, and protein content during synchronous and asynchronous growth of HeLa cells. Biophys. J. 9: 1377–1397; 1969.
Salzman, N. P. Systematic fluctuations in the cellular protein, RNA and DNA during growth of mammalian cell cultures. Biochim. Biophys. Acta 31: 158–163; 1959.
Spaggiare, S.; Wallach, M. J.; Tupper, J. T. Potassium transport in normal and transformed mouse 3T3 cells. J. Cell. Physiol. 89: 403–416; 1976.
Tsuboi, A.; Kurotsu, T.; Terasima, T. Changes in protein content per cell during growth of mouse L cells. Exp. Cell Res. 103: 257–261; 1976.
Volpe, P.; Eremenko-Volpe T. Quantitative studies on cell proteins in suspension cultures. Eur. J. Biochem. 12: 195–200; 1970.
Lowry, O. H.; Rosebrough, N. J.; Farr, A. L.; Randall, R. J. Protein measurement with the Folin phenol reagent. J. Biol. Chem. 193: 265–275; 1951.
Kletzien, R. F.; Pariza, M. W.; Becker, J. E.; Potter, V. R. A method using 3-0-methyl-d-glucose and phloretin for the determination of intracellular water space of cells in monolayer culture. Anal. Biochem. 68: 537–544; 1975.
Saier, M. H., Jr Growth and differentiated properties of an established kidney epithelial cell line (MDCK). Am. J. Physiol. 240: C106-C109; 1980.
Rindler, M. J.; McRoberts, J.; Saier, M. H., Jr. Na+, K+ co-transport in dog kidney epithelial cells (MDCK) (abstr.). J. Supramol. Struct 4: 86: 1980.
Parnes, J. R.; Isselbacher, K. J. Transport alterations in virus-transformed cells. Prog. Exp. Tumor Res. 22: 79–122; 1978.
Bakker-Grunwald, T. Effect of anions on potassium self-exchange in ascites tumor cells. Biochim. Biophys. Acta 513: 292–295; 1978.
Chipperfield, A. R. An effect of chloride on (Na+K) co-transport in red blood cells. Nature 286: 281–282; 1980.
Dunham, P. B.; Stewart, G. W.; Ellory, J. C. Chloride-activated passive potassium transport in human erythrocytes. Proc. Natl. Acad. Sci. USA 77: 1711–1715; 1980.
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This work was supported by USPHS Grant 5 R01 AM 21994-02 and NIH Grant AM 21506. S. E. was an Eli Lilly International Fellow and also supported by NATO and the Roussel-Uclaf Institute.
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Erlinger, S., Saier, M.H. Decrease in protein content and cell volume of cultured dog kidney epithelial cells during growth. In Vitro 18, 196–202 (1982). https://doi.org/10.1007/BF02618571
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DOI: https://doi.org/10.1007/BF02618571