Cell Migration pp 449-460 | Cite as

Beads on the Run: Beads as Alternative Tools for Chemotaxis Assays

Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 769)

Abstract

Cell migration is required for a wide variety of processes from bacteria seeking for food to correct patterning of neuronal networks. The ability to sense external cues is critical for cells to get directions and reach their goals. So far, studies on chemotaxis have mainly focused their attention on individual cells and therefore available tools are designed to monitor cell behavior at the single cell level. However, as collective cell migration is now widely accepted as a main mode of cell migration from development to cancer, the question of how chemotaxis is achieved has also to be asked on a bigger scale. Here, we present two chemotaxis assays suitable for single cells, cell sheets, and cell explants. Using a simple combination of heparin-coated beads and high vacuum silicone grease, these techniques can be adapted to a wide variety of culture conditions. They allow time-lapse study, high-resolution microscopy, and can be set up at no extra cost.

Key words

Chemotaxis Chemokine Growth factors Collective cell migration Cell culture Neural crest Mesoderm Growth cone Xenopus 

Notes

Acknowledgments

This investigation was supported by grants from MRC, BBSRC, and Wellcome Trust. We thank Rachel Moore for corrections and comments on the manuscript.

References

  1. 1.
    Serafini, T., Colamarino, S. A., Leonardo, E. D., Wang, H., Beddington, R., Skarnes, W. C., and Tessier-Lavigne, M. (1996) Netrin-1 is required for commissural axon guidance in the developing vertebrate nervous system, Cell 87, 1001–1014.PubMedCrossRefGoogle Scholar
  2. 2.
    Tessier-Lavigne, M., and Goodman, C. S. (1996) The molecular biology of axon guidance, Science 274, 1123–1133.PubMedCrossRefGoogle Scholar
  3. 3.
    Ming, G. L., Song, H. J., Berninger, B., Holt, C. E., Tessier-Lavigne, M., and Poo, M. M. (1997) cAMP-dependent growth cone guidance by netrin-1, Neuron 19, 1225–1235.PubMedCrossRefGoogle Scholar
  4. 4.
    Song, H. J., Ming, G. L., and Poo, M. M. (1997) cAMP-induced switching in turning direction of nerve growth cones, Nature 388, 275–279.PubMedCrossRefGoogle Scholar
  5. 5.
    Fernandis, A. Z., and Ganju, R. K. (2001) Slit: a roadblock for chemotaxis, Sci STKE 2001, pe1.Google Scholar
  6. 6.
    Boldajipour, B., Mahabaleshwar, H., Kardash, E., Reichman-Fried, M., Blaser, H., Minina, S., Wilson, D., Xu, Q., and Raz, E. (2008) Control of chemokine-guided cell migration by ligand sequestration, Cell 132, 463–473.PubMedCrossRefGoogle Scholar
  7. 7.
    Doitsidou, M., Reichman-Fried, M., Stebler, J., Koprunner, M., Dorries, J., Meyer, D., Esguerra, C. V., Leung, T., and Raz, E. (2002) Guidance of primordial germ cell migration by the chemokine SDF-1, Cell 111, 647–659.PubMedCrossRefGoogle Scholar
  8. 8.
    Richardson, B. E., and Lehmann, R. (2010) Mechanisms guiding primordial germ cell migration: strategies from different organisms, Nat Rev Mol Cell Biol 11, 37–49.PubMedCrossRefGoogle Scholar
  9. 9.
    Chen, A., Ganor, Y., Rahimipour, S., Ben-Aroya, N., Koch, Y., and Levite, M. (2002) The neuropeptides GnRH-II and GnRH-I are produced by human T cells and trigger laminin receptor gene expression, adhesion, chemotaxis and homing to specific organs, Nat Med 8, 1421–1426.PubMedCrossRefGoogle Scholar
  10. 10.
    Butcher, E. C., and Picker, L. J. (1996) Lymphocyte homing and homeostasis, Science 272, 60–66.PubMedCrossRefGoogle Scholar
  11. 11.
    Le, Y., Zhou, Y., Iribarren, P., and Wang, J. (2004) Chemokines and chemokine receptors: their manifold roles in homeostasis and disease, Cell Mol Immunol 1, 95–104.PubMedGoogle Scholar
  12. 12.
    Pals, S. T., de Gorter, D. J., and Spaargaren, M. (2007) Lymphoma dissemination: the other face of lymphocyte homing, Blood 110, 3102–3111.PubMedCrossRefGoogle Scholar
  13. 13.
    Boyden, S. (1962) The chemotactic effect of mixtures of antibody and antigen on polymorphonuclear leucocytes, J Exp Med 115, 453–466.PubMedCrossRefGoogle Scholar
  14. 14.
    Lauffenburger, D., Rothman, C., and Zigmond, S. H. (1983) Measurement of leukocyte motility and chemotaxis parameters with a linear under-agarose migration assay, J Immunol 131, 940–947.PubMedGoogle Scholar
  15. 15.
    Zicha, D., Dunn, G. A., and Brown, A. F. (1991) A new direct-viewing chemotaxis chamber, J Cell Sci 99 (Pt 4), 769–775.PubMedGoogle Scholar
  16. 16.
    Gundersen, R. W., and Barrett, J. N. (1979) Neuronal chemotaxis: chick dorsal-root axons turn toward high concentrations of nerve growth factor, Science 206, 1079–1080.PubMedCrossRefGoogle Scholar
  17. 17.
    Bianco, A., Poukkula, M., Cliffe, A., Mathieu, J., Luque, C. M., Fulga, T. A., and Rorth, P. (2007) Two distinct modes of guidance signalling during collective migration of border cells, Nature 448, 362–365.PubMedCrossRefGoogle Scholar
  18. 18.
    Deisboeck, T. S., and Couzin, I. D. (2009) Collective behavior in cancer cell populations, Bioessays 31, 190–197.PubMedCrossRefGoogle Scholar
  19. 19.
    Friedl, P., and Gilmour, D. (2009) Collective cell migration in morphogenesis, regeneration and cancer, Nat Rev Mol Cell Biol 10, 445–457.PubMedCrossRefGoogle Scholar
  20. 20.
    Rorth, P. (2009) Collective cell migration, Annu Rev Cell Dev Biol 25, 407–429.PubMedCrossRefGoogle Scholar
  21. 21.
    Theveneau, E., Marchant, L., Kuriyama, S., Gull, M., Moepps, B., Parsons, M., and Mayor, R. (2010) Collective chemotaxis requires contact-dependent cell polarity, Dev Cell 19, 39–53.PubMedCrossRefGoogle Scholar
  22. 22.
    Zigmond, S. H., and Hirsch, J. G. (1973) Leukocyte locomotion and chemotaxis. New methods for evaluation, and demonstration of a cell-derived chemotactic factor, J Exp Med 137, 387–410.Google Scholar
  23. 23.
    Orr, W., and Ward, P. A. (1978) Quantitation of leukotaxis in agarose by three different methods, J Immunol Methods 20, 95–107.PubMedCrossRefGoogle Scholar
  24. 24.
    Pujic, Z., Giacomantonio, C. E., Unni, D., Rosoff, W. J., and Goodhill, G. J. (2008) Analysis of the growth cone turning assay for studying axon guidance, J Neurosci Methods 170, 220–228.PubMedCrossRefGoogle Scholar
  25. 25.
    Xu, X., Brzostowski, J. A., and Jin, T. (2009) Monitoring dynamic GPCR signaling events using fluorescence microscopy, FRET imaging, and single-molecule imaging, Methods Mol Biol 571, 371–383.Google Scholar
  26. 26.
    Hynes, R. O. (2009) The extracellular matrix: not just pretty fibrils, Science 326, 1216–1219.PubMedCrossRefGoogle Scholar
  27. 27.
    He, L., and Niemeyer, B. (2003) A novel correlation for protein diffusion coefficients based on molecular weight and radius of gyration, Biotechnol Prog 19, 544–548.PubMedCrossRefGoogle Scholar
  28. 28.
    Young, M. E., Carroad, P. A., and Bell, R. L. (1980) Estimation of Diffusion-Coefficients of Proteins, Biotechnology and Bioengineering 22, 947–955.CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2011

Authors and Affiliations

  1. 1.Department of Cell and Developmental BiologyUniversity College LondonLondonUK

Personalised recommendations