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Methodological Aspects of Microelectrode Measurements in Cellular Spheroids

  • H. Acker
  • G. Holtermann
  • J. Carlsson
  • T. Nederman
Part of the Advances in Experimental Medicine and Biology book series (AEMB, volume 159)

Summary

Different types of oxygen microelectrodes have been tested in measurements on cellular spheroids. The shape of the oxygen gradients varied strongly depending on size and type of the spheroids. No significant differences in the results were obtained when different types of electrodes were applied. All measurements were made in a perfusion chamber. The shape of the gradients did not vary with time in the perfusion chamber. The reproducibility was found good in repeated measurements using the same spheroid. No mechanical or chemical disturbances were seen during the penetration of the spheroids. Changes in the medium flow rate through the chamber did not drastically change the shape of the oxygen gradients. Almost no convection could be seen at the bottom of the chamber close to the spheroids. The composition of the medium was found to be of importance. Lock’s solution containing glucose was found to be satisfactory. The potential signals in the double barrel electrodes allowed an accurate determination of the position when the electrode hit the spheroid surface. The information gained from microelectrode measurements in spheroids might be valuable for the understanding of effects of new tumor treatment modalities in which hypoxic cell sensitizers or high LET radiation are utilized.

Keywords

Methodological Aspect Oxygen Electrode Oxygen Gradient Perfusion Chamber Medium Flow Rate 
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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Reference

  1. 1.
    Acker, H., Carlsson, J. Measurements of potassium activities and membrane potentials in tumor cells. In: Progress in Enzyme and Ion-selective Electrode, D. W. Lubbers, H. Acker, R. P. Buck, G., Eisenman, M., Kessler, W., Simon (Eds) Springer, Berlin-Heidelberg-New York, 1981, pp 226 - 230.CrossRefGoogle Scholar
  2. 2.
    Acker, H., Dufau, E., Sylvester, D. Ionsensitive micro-electrodes with extra thin tip diameter for intracellular measurements. Pflügers Arch. 373, R 91, 1978b.Google Scholar
  3. 3.
    Acker, H., Sylvester, D., Dufau, E. and Durst, H. The Bitumen p02 electrode, a new method to manufacture p02 needle electrodes. In: Oxygen Transport to Tissue, Vol. III, I. A. Silver, M. Erecinska, H. I. Bicher (Eds.) Plenum Press, New York, 1978a, pp. 3 - 8.Google Scholar
  4. 4.
    Ackerman, L. W., Rosai, J. Surgical Pathology, C. W. Mosby Company, St. Louis 1974.Google Scholar
  5. 5.
    Baumgärtl, H., Lubbers, D. W., Platinum needle electrode for polarographic measurement of oxygen and hydrogen. In: Oxygen Supply. M. Kessler, D. F. Burley, L. C. Clark, Jr., D. W. Libbers, I. A. Silver, J. Strauss (Eds). Urban & Schwarzenberg, Munchen, 1973, pp. 130 - 136.Google Scholar
  6. 6.
    Carlsson, J. A proliferation gradient in three-dimensional colonies of cultured human glioma cells. Int. J. Cancer 20, 129-136A 1977.Google Scholar
  7. 7.
    Carlsson, J., Stalnacke, C. G., Acker, H., Haji-Karim, M.,Nilsson, S., Larssons, B. The influence of oxygen on proliferation and viability in cellular spheroids. Int. Rad. Onc. Biol. Phys. 5, 2011 - 2020, 1979.Google Scholar
  8. 8.
    Denekamp, J., Fowler, J. F. Cell proliferation kinetics and radiation therapy. Cancer, Vol. 6, F. F. Becker (Ed). Plenum Press, New York, 1977, pp. 105 - 107.Google Scholar
  9. 9.
    Dufau, E., Acker, H., Sylvester, D. Double-barrel ion-sensitive microelectrodes with extra thin tip diameters for intracellular measurements. Med. Prog. Technol. 7, 35 - 39, 1980.PubMedGoogle Scholar
  10. 10.
    Durand, R. E. Variable radiobiological response of spheroids. Radiat. Res. 81, 85 - 99, 1980.CrossRefGoogle Scholar
  11. 1l.
    Gray, L. H. Radiobiologic basis of oxygen as a modifying factor in radiation therapy. Amer. J. Roentgenol. Rad. Therap. and Nuclear Med. 85, 803 - 815, 1961.Google Scholar
  12. 12.
    Haji-Karim, M., Carlsson, J. Proliferation and viability in cellular spheroids of human origin. Cancer Res. 38, 1457 1464, 1970.Google Scholar
  13. 13.
    Hause, L. L., Pattilo, R. A., Sances, A., Mattingly, R. F. Cell surface coatings and membrane potentials of malignant and nonmalignant cells. Science 169, 601 - 603, 1970.PubMedCrossRefGoogle Scholar
  14. 14.
    Kallman, R. F. The phenomenon of reoxygenation and its implications for radiotherapy. Radiology 105, 135 - 142, 1972.PubMedGoogle Scholar
  15. 15.
    Litthrand, B., Revesz, L. The effect of oxygen on cellular survival and recovery after irradiation. Br. J. Radiol. 42, 914 - 924, 1969.CrossRefGoogle Scholar
  16. 16.
    Rubin, P., Casarett, G. W. Clinical Radiation Pathology, Vol. II, W. B. Saunders Company, Philadelphia 1968, pp. 934972.Google Scholar
  17. 17.
    Sutherland, R. M., Durand, R. E. Radiation response of multicell spheroids. An in vitro tumor model. Current Topics in Rad. Res. Quarterly Vol. 11, No. 1, 87 - 139, 1976.Google Scholar
  18. 18.
    Sutherland, R. M., McCredie, J. A., Inch, W. R. Growth of Multicell spheroids in tissue culture as a model of nodular carcinomas. J. Nat. Cancer Inst. 46, 113 - 120, 1971.PubMedGoogle Scholar
  19. 19.
    Tannock, I. The relation between cell proliferation and the vascular system in a transplanted mouse mammary tumor. Brit. J. Cancer 22, 258 - 273, 1968.PubMedCrossRefGoogle Scholar
  20. 20.
    Thonlinson, R. H., Gray, L. H. The histologic structure of sane human lung cancers and the possible implications for radiotherapy. Brit. J. Cancer 9, 539 - 549, 1955.CrossRefGoogle Scholar
  21. 21.
    Vaupel, P., Frinak, S. and Bicher, H. I. Heterogeneous p02 and pH distributions in C3H-mouse mammary carcinoma. Presented at the 4th annual meeting of the Gesellschaft für Mikrozirkulation, Mainz, 1979.Google Scholar
  22. 22.
    Yuhas, J. M., Li, A. P., Martinez, A. O., Ladman, A. J. A simplified method for production and growth of multi-cellular tumor spheroids. Cancer Res. 37, 3639 - 3643, 1977.PubMedGoogle Scholar

Copyright information

© Plenum Press, New York 1983

Authors and Affiliations

  • H. Acker
    • 1
  • G. Holtermann
    • 1
  • J. Carlsson
    • 2
  • T. Nederman
    • 3
  1. 1.Max-Planck-Institut fof SystemphysiologyDortmund 1Germany
  2. 2.Department of RadiobiologyNational Defense Research InstituteUmeåSweden
  3. 3.Department of Physical BiologyGustaf Werner InstituteUppsalaSweden

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