Summary
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1.
Mouse LS cells grow in completely mixed steady-state continuous suspension (“chemostat”) culture in defined medium.
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2.
The steady-state concentration of cells is maximal at a dilution rate of 0.30 to 0.35 day−1.
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3.
Glucose can act as the limiting substrate for LS cells under chemostat conditions.
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4.
The glucose oxidation rate per cell does not vary with dilution rate.
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5.
Maintenance energy is 19 picomoles of ATP per cell per day. Growth energy is 22 picomoles of ATP per cell.
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6.
Slowly growing cells contain more protein and less RNA per cell than rapidly growing cells.
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7.
The “efficiency” of protein synthesis decreases in slowly growing cells, in which a lower proportion of ribosomes is present in the form of polysomes or ribosomal subunits.
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8.
Newly-made 18S RNA appears early in the cytoplasm of rapidly growing cells, but is greatly delayed in slowly growing cells.
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9.
Pulsed additions of a limiting substrate to steady-state populations may lead to synchronized cells that have a controlled interdivision time. Hence chemostat cultures may be used to investigate the interdependence of events in the cell cycle.
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References
Pirt, S. J. 1972. Prospects and problems in continuous fow culture of micro-organisms. J. Appl. Chem. Biotechnol. 22: 55–64.
Cohen, E. P., and H. Eagle. 1961. A simfied chemostat for the growth of mammalian cells: characteristics of cell growth in continuous culture. J. Exp. Med. 13: 467–474.
Sinclair, R., R. A. Reid, and P. Mitchell. 1963. Culture of strain L cells in suspension: replacement of polymer by traces of trypsin in a defined medium. Nature 197: 982–984.
Sinclair, R. 1966. Steady-state suspension culture and metabolism of strain L mouse cells in simple defined medium. Exp. Cell Res. 41: 20–31.
Pirt, S. J., and D. S. Callow. 1964. Continuous flow culture of the ERK and L types of mammalian cells. Exp. Cell Res. 33: 413–421.
Griffiths, J. B., and S. J. Pirt. 1967. The uptake of amino acids by mouse cells (strain L S) during growth in batch culture and chemostat culture: the influence of cell growth rate. Proc. R. Soc. B. 168: 421–438.
Moser, H., and G. Vecchio. 1967. The production of stable steady-states in mouse ascites mast cell cultures maintained in a chemostat. Experientia 23: 120–123.
Peraino, C., S. Bacchetti, and W. J. Eisler. 1970. Automated continuous culture of mammalian cells in suspension. Science 169: 204–205.
Herbert, D., R. Ellsworth, and R. C. Telling. 1956. The continuous culture of bacteria; a theoretical and experimental study. J. Gen. Microbiol. 14: 601–622.
Birch, J. R., and S. J. Pirt. 1970. Improvements in a chemically defined medium for the growth of mouse cells (strain L S) in suspension. J. Cell Sci. 7: 661–670.
Kubitschek, H. E. 1971.Introduction to Research with Continuous Cultures. Prentice-Hall, Inc., Englewood-Cliffs, N. J., Chap. 4.
Williams, F. M. 1967 A model of cell growth dynamics. J. Theor. Biol. 15: 190–207.
Pirt, S. J. 1965 The maintenance energy of bacteria in growing cultures. Proc. R. Soc. B. 163: 224–231.
Wase, D. A. J., and J. S. Hough 1966. Continuous culture of yeast on phenol. J. Gen Microbiol. 42: 13–23.
Kilburn, D. G., M. D. Lilly and F. C. Webb. 1969. The energetics of mammalian cell growth. J. Cell Sci. 4: 645–654.
Sinclair, C. G., and H. H. Topiwala. 1970. Model for continuous culture which considers the viability concept. Biotechnol. Bioeng. 12: 1069–1079.
Kilburn, D. G., M. D. Lilly, D. A. Self, and F. C. Webb. 1969. The effect of dissolved oxygen partial pressure on the growth and carbohydrate metabolism of mouse LS cells. J. Cell Sci. 4: 25–37.
Radlett, P. J., R. C. Telling, J. P. Whiteside, and M. A. Maskell. 1972. The supply of oxygen to submerged cultures of BHK21 cells. Biotechnol. Bioeng. 14: 437–445.
Barton, M. E. 1971. Effect of pH on the growth cycle of HeLa cells in batch suspension culture without oxygen control. Biotechnol. Bioeng. 13: 471–492.
Glinos, A. D., R. J. Werrlein, and N. M. Papadopoulos. 1965. Constitution, viability and lactate dehydrogenase in stationary-phase L-cell suspension cultures. Science 150: 350–353.
Self, D. A., D. G. Kilburn, and M. D. Lilly. 1968. The influence of dissolved oxygen partial pressure on the level of various enzymes in mouse LS cells. Biotechnol. Biong. 10: 815–828.
Criss, W. E. 1973. Control of the adenylate charge in the Morris “minimal-deviation” hepatomas. Cancer Res. 33: 51–56.
Ecker, T. E., and M. Schaechter. 1963. Ribosome content and the rate of growth ofSalmonella typhimurium. Biochim. Biophys. Acta 76: 275–279.
Sykes, J., and T. W. Young. 1968. Studies on ribosomes and ribonucleic acids ofAerobacter aerogenes grown at different rates in a carbon-limited continuous culture. Biochim. Biophys. Acta 169: 103–116.
Hogan, B. L. M., and A. Korner. 1968. Ribosomal subunits of Landschutz ascites cells during changes in polysomal distribution. Biochim. Biophys. Acta 169: 129–138.
Joklik, W. K., and Y. Becker. 1965. Studies on the genesis of polyribosomes, II. The association of nascent messenger RNA with the 40 S subribosomal particles. J. Mol. Biol. 13: 511–520.
Koch, A. L. 1971. The adaptive responses ofEscherichia coli to a feast and famine existence. Adv. Microbiol. Physiol. 6: 147–217.
Daskal, I. 1971. Ph.D. Thesis, McGill University, Montreal.
Hansche, P. F. 1969. A theoretical basis for the entrainment of chemostat populations. J. Theor. Biol. 24: 335–350.
Franke, E. K. 1970. A mathematical model of synchronized periodic growth of cell populations. J. Theor. Biol. 26: 373–382.
Dawson, P. S. S. 1972. Continuously synchronized growth. In: A. C. R. Dean, S. J. Pirt, and D. W. Tempest (Eds.)Environmental Control of Cell Synthesis and Function. Academic Press, Inc. London, pp. 79–103.
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Sinclair, R. Response of mammalian cells to controlled growth rates in steady-state continuous culture. In Vitro 10, 295–305 (1974). https://doi.org/10.1007/BF02615311
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DOI: https://doi.org/10.1007/BF02615311