Abstract
We present a microfabricated chamber designed for visualising and quantifying the chemotaxis of slow-migrating adherent mammalian cells such as cancer and endothelial cells. Most of the existing solutions for the investigation of chemotaxis are limited to fast migrating cells such as leukocytes or Dictyostelium discoideum. Here, we describe the details of an assay using the μ-Slide Chemotaxis to investigate the chemotactic response of human umbilical vein endothelial cells to a gradient of human vascular endothelial growth factor 165. In combination with phase contrast video microscopy and cell tracking, the trajectories of all single cells migrating in temporally stable gradients are derived. The resulting migration data are displayed and analysed in detail by several different parameters for quantifying chemotaxis. We found that with this tool the potential of chemoattractants to migration of mammalian cells as well as the impact of inhibitors to chemotaxis and migration can be evaluated.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Cazes, A., Galaup, A., Chomel, C., Bignon, M., Brechot, N., Le Jan, S., Weber, H., Corvol, P., Muller, L., Germain, S., and Monnot, C. (2006) Extracellular Matrix-Bound Angiopoietin-Like 4 Inhibits Endothelial Cell Adhesion, Migration, and Sprouting and Alters Actin Cytoskeleton, Circ Res 99, 1207–1215.
Harris, M. P., Kim, E., Weidow, B., Wikswo, J. P., and Quaranta, V. (2008) Migration of isogenic cell lines quantified by dynamic multivariate analysis of single-cell motility, Cell Adh Migr 2, 127–136.
Lin, F., and Butcher, E. C. (2006) T cell chemotaxis in a simple microfluidic device, Lab Chip 6, 1462–1469.
Fisher, P. R., Merkl, R., and Gerisch, G. (1989) Quantitative analysis of cell motility and chemotaxis in Dictyostelium discoideum by using an image processing system and a novel chemotaxis chamber providing stationary chemical gradients, J Cell Biol 108, 973–984.
Schneider, L., Cammer, M., Lehman, J., Nielsen, S. K., Guerra, C. F., Veland, I. R., Stock, C., Hoffmann, E. K., Yoder, B. K., Schwab, A., Satir, P., and Christensen, S. T. (2010) Directional cell migration and chemotaxis in wound healing response to PDGF-AA are coordinated by the primary cilium in fibroblasts, Cell Physiol Biochem 25, 279–292.
Lamalice, L., Le Boeuf, F., and Huot, J. (2007) Endothelial cell migration during angiogenesis, Circ Res 100, 782–794.
Cooper, C. R., and Pienta, K. J. (2000) Cell adhesion and chemotaxis in prostate cancer metastasis to bone: a minireview, Prostate Cancer Prostatic Dis 3, 6–12.
Zigmond, S. H. (1988) Orientation chamber in chemotaxis, Methods Enzymol 162, 65–72.
Boyden, S. (1962) The chemotactic effect of mixtures of antibody and antigen on polymorphonuclear leucocytes, J Exp Med 115, 453–466.
Eiseler, T., Doppler, H., Yan, I. K., Kitatani, K., Mizuno, K., and Storz, P. (2009) Protein kinase D1 regulates cofilin-mediated F-actin reorganization and cell motility through slingshot, Nat Cell Biol 11, 545–556.
Molina-Ortiz, I., Bartolome, R. A., Hernandez-Varas, P., Colo, G. P., and Teixido, J. (2009) Overexpression of E-cadherin on melanoma cells inhibits chemokine-promoted invasion involving p190RhoGAP/p120ctn-dependent inactivation of RhoA, J Biol Chem 284, 15147–15157.
Rothmeier, A. S., Ischenko, I., Joore, J., Garczarczyk, D., Furst, R., Bruns, C. J., Vollmar, A. M., and Zahler, S. (2009) Investigation of the marine compound spongistatin 1 links the inhibition of PKCalpha translocation to nonmitotic effects of tubulin antagonism in angiogenesis, FASEB J 23, 1127–1137.
Zigmond, S. H., Foxman, E. F., and Segall, J. E. (2001) Chemotaxis assays for eukaryotic cells, Curr Protoc Cell Biol Chapter 12, Unit 12 11.
Foxman, E. F., Kunkel, E. J., and Butcher, E. C. (1999) Integrating conflicting chemotactic signals. The role of memory in leukocyte navigation, J Cell Biol 147, 577–588.
Fisher, N. I. (1993) Statistical Analysis of Circular Data, Cambridge University Press, New York.
Acknowledgments
This work was supported by the BMBF grants “Schwerpunktprogramm Mikrosystemtechnik (BioMST)” and “Optische Technologien (Biophotonik).”
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2011 Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Zantl, R., Horn, E. (2011). Chemotaxis of Slow Migrating Mammalian Cells Analysed by Video Microscopy. In: Wells, C., Parsons, M. (eds) Cell Migration. Methods in Molecular Biology, vol 769. Humana Press. https://doi.org/10.1007/978-1-61779-207-6_13
Download citation
DOI: https://doi.org/10.1007/978-1-61779-207-6_13
Published:
Publisher Name: Humana Press
Print ISBN: 978-1-61779-206-9
Online ISBN: 978-1-61779-207-6
eBook Packages: Springer Protocols