Abstract
The Boyden chamber assay, originally introduced by Boyden for the analysis of leukocyte chemotaxis, is based on a chamber of two medium-filled compartments separated by a microporous membrane. In general, cells are placed in the upper compartment and are allowed to migrate through the pores of the membrane into the lower compartment, in which chemotactic agents are present. After an appropriate incubation time, the membrane between the two compartments is fixed and stained, and the number of cells that have migrated to the lower side of the membrane is determined. Therefore, the Boyden chamber-based cell migration assay has also been called filter membrane migration assay, trans-well migration assay, or chemotaxis assay. A number of different Boyden chamber devices are available commercially. The method described in this chapter is intended specifically for measuring the migration of Madin-Darby canine kidney cells using a 48-well chamber from Neuro Probe, Inc.
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References
Boyden, S. V. (1962) The chemotactic effect of mixtures of antibody and antigen on polymorphonuclear leucocytes. J. Exp. Med. 115, 453–466.
Carter, S. B. (1967) Heptotaxis and the mechanism of cell motility. Nature 213, 256–260.
Reiske, H. R., Kao, S-C., Cary, L.A., Guan, J-L., Lai, J-F., and Chen, H-C. (1999) Requirement of phosphatidylinositol 3-kinase in focal adhesion kinase-promoted cell migration. J. Biol. Chem. 274, 12,361–12,366.
Liang, C-C. and Chen, H-C. (2001) Sustained activation of extracellular signal-regulated kinase stimulated by hepatocyte growth factor leads to integrin (β2 expression that is involved in cell scattering. J. Biol. Chem. 276, 21,146–21,152.
Cary, L. A., Chang, J. F., and Guan, J-L. (1996) Stimulation of cell migration by overexpression of focal adhesion kinase and its association with Src and Fyn. J. Cell Sci. 109, 1787–1794.
Sander E. E., van Delft, S., ten Klooster, J. P., Reid, T., van der Kammen, R. A., Michiels, F., et al. (1998) Matrix-dependent Tiam1/rac signaling in epithelial cells promotes either cell-cell adhesion or cell migration and is regulated by phosphatidylinositol 3-kinase. J. Cell Biol. 143, 1385–1398.
Royal, I., Lamarche-Vane, N., Lamorte, L., Kaibuchi, K., and Park, M. (2000) Activation of Cdc42, Rac, PAK, and Rho-kinase in response to hepatocyte growth factor differentially regulates epithelial cell colony spreading and dissociation. Mol. Biol. Cell 11, 1709–1725.
Ren, X. D., Kiosses, W. B., Sieg, D. J., Otey, C. A., Schlaepfer, D. D., and Schwartz, M. A. (2000) Focal adhesion kinase suppresses Rho activity to promote focal adhesion turnover. J. Cell. Sci. 113, 3673–3678.
Chen, B-H., Tzen, J. T. C., Bresnick, A. R., and Chen, H-C. (2002) Roles of Rho-associated kinase and myosin light chain kinase in morphological and migratory defects of focal adhesion kinase-null cells. J. Biol. Chem. 277, 33,857–33,863.
Lai, J-F., Kao, S-C., Jiang, S-T., Tang, M-J., Chan, P-C., and Chen, H-C. (2000) Involvement of focal adhesion kinase in hepatocyte growth factor-induced scatter of Madin-Darby canine kidney cells. J. Biol. Chem. 275, 7474–7480.
Sieg, D. J., Hauck, C. R., and Schlaepfer D. D. (1999) Required role of focal adhesion kinase (FAK) for integrin-stimulated cell migration. J. Cell Sci. 112, 2677–2691.
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© 2005 Humana Press Inc., Totowa, NJ
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Chen, HC. (2005). Boyden Chamber Assay. In: Guan, JL. (eds) Cell Migration. Methods in Molecular Biology™, vol 294. Humana Press. https://doi.org/10.1385/1-59259-860-9:015
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DOI: https://doi.org/10.1385/1-59259-860-9:015
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