Advertisement

The Journal of Membrane Biology

, Volume 1, Issue 1, pp 109–124 | Cite as

Intercellular adhesion

I. A quantitative assay for measuring the rate of adhesion
  • Charles W. Orr
  • Saul Roseman
Article

Summary

A quantitative procedure for determining the early kinetics of cell aggregation (adhesion) is described. The cells used for this study were obtained by dissociation of 8-day-old embryonic chicken neural retina with crude trypsin. The method is based on determining the decrease in single cells in an aggregating population with the Coulter electronic particle counter. A variety of experiments show that the method is reproducible and capable of detecting relatively small changes in the rate of aggregation. Using a number of criteria, the loss of single cells from the population with increasing time of incubation was shown to result from the formation of aggregates, and not from other phenomena such as cell death or changes in cell permeability. The intercellular adhesions formed under these conditions were stable to mechanical shear and to ethylenediaminetetraacetate, and were partially resistant to crude trypsin. The logarithm10 of the number of single cells in the population was found to be directly related to the time of incubation. The slope of the resultant straight lines could be used as a measure of the rate of aggregation. No lag in aggregation was demonstrable under the standard assay conditions. the rate was affected by the initial cell density, speed of rotation during aggregation, temperature, and by Ca2+ and Mg2+. It was not affected by inhibitors of protein synthesis, metabolic inhibitors, ATP, ADP, cyclic-AMP, or horse serum at 37 °C. The quantitative method for determining the initial rate of adhesion should be applicable to studies on the chemistry of this process.

Keywords

Retina Single Cell Horse Serum Cell Aggregation Intercellular Adhesion 
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.

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. 1.
    Abercrombie, M., and E. J. Ambrose. 1962. The surface properties of cancer cells: A review.Cancer Res. 22:525.PubMedGoogle Scholar
  2. 2.
    Ball, W. D. 1963. A quantitative assessment of mouse thymus differentiation.Exp. Cell Res. 31:82.PubMedGoogle Scholar
  3. 3.
    — 1966. Aggregation of dissociated embryonic chick cells at 3°.Nature 210:1075.PubMedGoogle Scholar
  4. 4.
    Cook, G. M. W., D. H. Heard, and G. V. F. Seaman. 1960. A sialomucopeptide liberated by trypsin from the human erythrocyte.Nature 188:1011.PubMedGoogle Scholar
  5. 5.
    Curtis, A. S. G. 1962. Cell contact and adhesion.Biol. Rev. 37:82.PubMedGoogle Scholar
  6. 6.
    — 1963. The effect of pH and temperature on cell aggregation.Nature 200:1235.Google Scholar
  7. 7.
    —, and M. F. Greaves. 1965. The inhibition of aggregation by a pure serum protein.J. Embryol. 13:309.PubMedGoogle Scholar
  8. 8.
    Dolan, m. F. 1965. Viability assays — a critique.Fed. Proc. 24:5.PubMedGoogle Scholar
  9. 9.
    Jones, B. M. 1966. A unifying hypothesis of cell adhesion.Nature 212:362.PubMedGoogle Scholar
  10. 10.
    Kemp, R. B., B. M. Jones, I. Cunningham, and M. C. M. Jones. 1967. Quantitative investigation on the effect of puromycin on the aggregation of trypsin — and versene —dissociated chick fibroblast cells.J. Cell Sci. 2:323.PubMedGoogle Scholar
  11. 11.
    Knight, V. A., B. M. Jones, and P. C. T. Jones. 1966. Inhibition of aggregation of dissociated embryo chick fibroblast cells by adenosine triphosphate.Nature 210:1008.PubMedGoogle Scholar
  12. 12.
    Langley, D. K., and E. J. Ambrose. 1964. Isolation of a mucopeptide from the surface of Ehrlich ascites tumour cells.Nature 204:53.PubMedGoogle Scholar
  13. 13.
    Lillien, J. E. 1968. Specific enhancement of cell aggregationin vitro.Develop. Biol. 17:657.PubMedGoogle Scholar
  14. 14.
    Moscona, A. 1961. Rotation-mediated histogenic aggregation of dissociated cells.Exp. Cell Res. 22:455.PubMedGoogle Scholar
  15. 15.
    Moscona, A. A. 1963. Studies on cell aggregation: demostration of materials with a selective cell-binding activity.Proc. Nat. Acad. Sci., Wash. 49:742.Google Scholar
  16. 16.
    —, and M. H. Moscona. 1966. Aggregation of embryonic cells in a serum-free medium and its inhibition at suboptimal temperatures.Expl. Cell Res. 41:687.Google Scholar
  17. 17.
    Moscona, M. H., and A. A. Moscona. 1963. Inhibition of adhesiveness of dissociated cells by inhibitors of protein and RNA synthesis.Science 142:1070.PubMedGoogle Scholar
  18. 18.
    Orr, C. W. M., and S. Roseman. 1969. Intercellular adhesion. II. The purification and properties of a horse serum protein that promotes neural retina cell aggregation.J. Membrane Biol. 1:125.Google Scholar
  19. 19.
    Overbeek, J. Th. G. 1952. Chapter VII.In Colloid Science, Vol. I. H. R. Kruyt, editor. Elsevier Publishing Co., Amsterdam.Google Scholar
  20. 20.
    Overton, J. 1969. A fibrillar intercellular material between reaggregating embryonic chick cells.Expl. Cell Res. 40:136.Google Scholar
  21. 21.
    Rodman, N. F., and R. G. Mason. 1967. Platelet-platelet interaction: relationship to homeostasis and thrombosis.Fed. Proc. 26:95.PubMedGoogle Scholar
  22. 22.
    Roth, S. A., and J. A. Weston. 1967. The measurement of intercellular adhesion.Proc. Nat. Acad. Sci., Wash. 58:974.Google Scholar
  23. 23.
    Steinberg, M. S. 1958. On the chemical bonds between animal cells: a mechanism for type specific association.Am. Naturalist 92:65.Google Scholar
  24. 24.
    — 1962. The role of temperature in the control of aggregation of dissociated embryonic cells.Expl. Cell Res. 28:1.Google Scholar
  25. 25.
    — 1962. On the mechanism of tissue reconstruction by dissociated cells — 1. Population kinetics, differential adhesiveness and the absence of directed migration.Proc. Nat. Acad. Sci., Wash. 48:1577.Google Scholar
  26. 26.
    — 1963. “ECM”; its nature, origin and function in cell aggregation.Expl. Cell Res. 30:257.Google Scholar
  27. 27.
    Townes, P. L., and J. Holtfreter. 1955. Directed movements and selective adhesion of embryonic amphibian cells.J. Exp. Zool. 128:53.Google Scholar
  28. 28.
    Trinkaus, J. P. 1965. Mechanism of morphogenetic movements.In Organogenesis. R. L. DeHaan and H. Ursprung, editors. p. 55. Holt, Rinehart & Winston, Inc., New York.Google Scholar
  29. 29.
    Weiss, L., and E. Mayhew. 1967. The cell periphery.New Engl. J. Med. 276:1354.PubMedGoogle Scholar
  30. 30.
    Weyesse, A. W., and W. S. Burgess. 1906. Histogenesis of the retina.Am. Naturalist 40:611.Google Scholar

Copyright information

© Springer-Verlag New York Inc. 1969

Authors and Affiliations

  • Charles W. Orr
    • 1
    • 2
  • Saul Roseman
    • 1
    • 2
  1. 1.McCollum-Pratt InstituteThe Johns Hopkins UniversityBaltimore
  2. 2.Department of BiologyThe Johns Hopkins UniversityBaltimore

Personalised recommendations