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In Vitro

, Volume 10, Issue 5–6, pp 295–305 | Cite as

Response of mammalian cells to controlled growth rates in steady-state continuous culture

  • R. Sinclair
Article

Summary

  1. 1.

    Mouse LS cells grow in completely mixed steady-state continuous suspension (“chemostat”) culture in defined medium.

     
  2. 2.

    The steady-state concentration of cells is maximal at a dilution rate of 0.30 to 0.35 day−1.

     
  3. 3.

    Glucose can act as the limiting substrate for LS cells under chemostat conditions.

     
  4. 4.

    The glucose oxidation rate per cell does not vary with dilution rate.

     
  5. 5.

    Maintenance energy is 19 picomoles of ATP per cell per day. Growth energy is 22 picomoles of ATP per cell.

     
  6. 6.

    Slowly growing cells contain more protein and less RNA per cell than rapidly growing cells.

     
  7. 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.

     
  8. 8.

    Newly-made 18S RNA appears early in the cytoplasm of rapidly growing cells, but is greatly delayed in slowly growing cells.

     
  9. 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.

     

Keywords

Dilution Rate Continuous Culture Ribosomal Subunit Dilution Ratio Chemostat Culture 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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References

  1. 1.
    Pirt, S. J. 1972. Prospects and problems in continuous fow culture of micro-organisms. J. Appl. Chem. Biotechnol. 22: 55–64.Google Scholar
  2. 2.
    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.CrossRefGoogle Scholar
  3. 3.
    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.PubMedCrossRefGoogle Scholar
  4. 4.
    Sinclair, R. 1966. Steady-state suspension culture and metabolism of strain L mouse cells in simple defined medium. Exp. Cell Res. 41: 20–31.PubMedCrossRefGoogle Scholar
  5. 5.
    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.PubMedCrossRefGoogle Scholar
  6. 6.
    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.Google Scholar
  7. 7.
    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.PubMedCrossRefGoogle Scholar
  8. 8.
    Peraino, C., S. Bacchetti, and W. J. Eisler. 1970. Automated continuous culture of mammalian cells in suspension. Science 169: 204–205.PubMedCrossRefGoogle Scholar
  9. 9.
    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.PubMedGoogle Scholar
  10. 10.
    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.PubMedGoogle Scholar
  11. 11.
    Kubitschek, H. E. 1971.Introduction to Research with Continuous Cultures. Prentice-Hall, Inc., Englewood-Cliffs, N. J., Chap. 4.Google Scholar
  12. 12.
    Williams, F. M. 1967 A model of cell growth dynamics. J. Theor. Biol. 15: 190–207.PubMedCrossRefGoogle Scholar
  13. 13.
    Pirt, S. J. 1965 The maintenance energy of bacteria in growing cultures. Proc. R. Soc. B. 163: 224–231.Google Scholar
  14. 14.
    Wase, D. A. J., and J. S. Hough 1966. Continuous culture of yeast on phenol. J. Gen Microbiol. 42: 13–23.PubMedGoogle Scholar
  15. 15.
    Kilburn, D. G., M. D. Lilly and F. C. Webb. 1969. The energetics of mammalian cell growth. J. Cell Sci. 4: 645–654.PubMedGoogle Scholar
  16. 16.
    Sinclair, C. G., and H. H. Topiwala. 1970. Model for continuous culture which considers the viability concept. Biotechnol. Bioeng. 12: 1069–1079.PubMedCrossRefGoogle Scholar
  17. 17.
    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.PubMedGoogle Scholar
  18. 18.
    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.PubMedCrossRefGoogle Scholar
  19. 19.
    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.PubMedCrossRefGoogle Scholar
  20. 20.
    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.PubMedCrossRefGoogle Scholar
  21. 21.
    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.CrossRefGoogle Scholar
  22. 22.
    Criss, W. E. 1973. Control of the adenylate charge in the Morris “minimal-deviation” hepatomas. Cancer Res. 33: 51–56.PubMedGoogle Scholar
  23. 23.
    Ecker, T. E., and M. Schaechter. 1963. Ribosome content and the rate of growth ofSalmonella typhimurium. Biochim. Biophys. Acta 76: 275–279.PubMedCrossRefGoogle Scholar
  24. 24.
    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.PubMedGoogle Scholar
  25. 25.
    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.PubMedGoogle Scholar
  26. 26.
    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.PubMedCrossRefGoogle Scholar
  27. 27.
    Koch, A. L. 1971. The adaptive responses ofEscherichia coli to a feast and famine existence. Adv. Microbiol. Physiol. 6: 147–217.CrossRefGoogle Scholar
  28. 28.
    Daskal, I. 1971. Ph.D. Thesis, McGill University, Montreal.Google Scholar
  29. 29.
    Hansche, P. F. 1969. A theoretical basis for the entrainment of chemostat populations. J. Theor. Biol. 24: 335–350.PubMedCrossRefGoogle Scholar
  30. 30.
    Franke, E. K. 1970. A mathematical model of synchronized periodic growth of cell populations. J. Theor. Biol. 26: 373–382.PubMedCrossRefGoogle Scholar
  31. 31.
    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.Google Scholar

Copyright information

© Tissue Culture Association 1975

Authors and Affiliations

  • R. Sinclair
    • 1
  1. 1.Dept. of BiologyMcGill UniversityMontrealCanada

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