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Cell Migration Analyses Within Fibroblast-Derived 3-D Matrices

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Cell Migration

Part of the book series: Methods in Molecular Biology™ ((MIMB,volume 294))

Abstract

Research in cell biology often is based in tissue culturing cells on artificial substrates, such as plastic or glass. These artificial conditions are prone to distorting findings by persuading cells to adjust to artificial flat rigid surfaces. In contrast, the natural substrate for most cells in living organisms is the extracellular matrix (ECM), which is three-dimensional, complex, and dynamic in its molecular composition, and variable in pliability. Here we show how to measure the rates and directionality of fibroblasts motility within physiologically relevant in vivo-like cell-derived three-dimensional matrices.

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References

  1. Elsdale, T. and Bard, J. (1972) Collagen substrata for studies on cell behavior. J. Cell Biol. 54, 626–637.

    Article  PubMed  CAS  Google Scholar 

  2. Elsdale, T. and Bard, J. (1972) Cellular interactions in mass cultures of human diploid fibroblasts. Nature 236, 152–155.

    Article  PubMed  CAS  Google Scholar 

  3. Elsdale, T. and Bard, J. (1974) Cellular interactions in morphogenesis of epithelial mesenchymal systems. J. Cell Biol. 63, 343–349.

    Article  PubMed  CAS  Google Scholar 

  4. Friedl, P. and Brocker, E. B. (2002) Tcr triggering on the move: Diversity of t-cell interactions with antigen-presenting cells. Immunol. Rev. 186, 83–89.

    Article  PubMed  CAS  Google Scholar 

  5. Cukierman, E., Pankov, R., Stevens, D. R., and Yamada, K. M. (2001) Taking cell-matrix adhesions to the third dimension. Science 294, 1708–1712.

    Article  PubMed  CAS  Google Scholar 

  6. Streuli, C. (1999) Extracellular matrix remodelling and cellular differentiation. Curr. Opin. Cell Biol. 11, 634–640.

    Article  PubMed  CAS  Google Scholar 

  7. Badylak, S. (2002) The extracellular matrix as a scaffold for tissue reconstruction. Semin. Cell Dev. Biol. 13, 377–383.

    Article  PubMed  CAS  Google Scholar 

  8. Buck, C. A. and Horwitz, A. F. (1987) Cell surface receptors for extracellular matrix molecules. Annu. Rev. Cell Biol. 3, 179–205.

    Article  PubMed  CAS  Google Scholar 

  9. Hay, E. D. (1991) Cell Biology of Extracellular Matrix, 2nd Ed., Plenum Press, New York, NY.

    Google Scholar 

  10. Hynes, R. O. (1999) Cell adhesion: Old and new questions. Trends Cell Biol. 9, M33–M77.

    Article  PubMed  CAS  Google Scholar 

  11. Geiger, B., Bershadsky, A., Pankov, R., and Yamada, K. M. (2001) Transmembrane crosstalk between the extracellular matrix and the cytoskeleton. Nat. Rev. Mol. Cell Biol. 2, 793–805.

    Article  PubMed  CAS  Google Scholar 

  12. Walpita, D. and Hay, E. (2002) Studying actin-dependent processes in tissue culture. Nat. Rev. Mol. Cell Biol. 3, 137–141.

    Article  PubMed  CAS  Google Scholar 

  13. Bissell, M. J., Radisky, D. C, Rizki, A., Weaver, V. M., and Petersen, O. W. (2002) The organizing principle: Microenvironmental influences in the normal and malignant breast. Differentiation 70, 537–546.

    Article  PubMed  Google Scholar 

  14. Miranti, C. K. and Brugge, J. S. (2002) Sensing the environment: A historical perspective on integrin signal transduction. Nat. Cell Biol. 4, E83–E90.

    Article  PubMed  CAS  Google Scholar 

  15. Bosman, F. T. and Stamenkovic, I. (2003) Functional structure and composition of the extracellular matrix. J. Pathol. 200, 423–428.

    Article  PubMed  CAS  Google Scholar 

  16. Clegg, D. O., Wingerd, K. L., Hikita, S. T., and Tolhurst, E. C. (2003) Integrins in the development, function and dysfunction of the nervous system. Front. Biosci. 8, D723–D750.

    Article  PubMed  CAS  Google Scholar 

  17. Schmeichel, K. L. and Bissell, M. J. (2003) Modeling tissue-specific signaling and organ function in three dimensions. J. Cell Sci. 116, 2377–2388.

    Article  PubMed  CAS  Google Scholar 

  18. Wakatsuki, T. and Elson, E. L. (2003) Reciprocal interactions between cells and extracellular matrix during remodeling of tissue constructs. Biophys. Chem. 100, 593–605.

    Article  PubMed  CAS  Google Scholar 

  19. Yamada, K. M., Pankov, R., and Cukierman, E. (2003) Dimensions and dynamics in integrin function. Braz. J. Med. Biol. Res. 36, 959–966.

    Article  PubMed  CAS  Google Scholar 

  20. Kunz-Schughart, L. A., Heyder, P., Schroeder, J., and Knuechel, R. (2001) A heterologous 3-D coculture model of breast tumor cells and fibroblasts to study tumor-associated fibroblast differentiation. Exp. Cell Res. 266, 74–86.

    Article  PubMed  CAS  Google Scholar 

  21. Olumi, A. F., Grossfeld, G. D., Hayward, S. W., Carroll, P. R., Tlsty, T. D., and Cunha, G. R. (1999) Carcinoma-associated fibroblasts direct tumor progression of initiated human prostatic epithelium. Cancer Res. 59, 5002–5011.

    PubMed  CAS  Google Scholar 

  22. Akiyama, S. K. (1999) Purification of fibronectin, in Current Protocols in Cell Biology (Bonifacino, J. S., Dasso, M., Lippincott-Schwartz, J., Harford, J. B., and Yamada, K. M., eds.) John K. Wiley&Sons, New York, NY, Vol. 1, pp. 10.15.11–10.15.13.

    Google Scholar 

  23. Cukierman, E. (2002) Preparation of extracellular matrices produced by cultured fibroblasts, in Current Protocols in Cell Biology (Bonifacino, J. S, Dasso, M., Lippincott-Schwartz, J., Harford, J. B., and Yamada, K. M., eds.) John K. Wiley &Sons, New York, NY, Vol. 1, pp. 10.19.11–10.19.14.

    Google Scholar 

  24. Cukierman, E., Pankov, R., and Yamada, K. M. (2002) Cell interactions with three-dimensional matrices. Curr. Opin. Cell Biol. 14, 633–640.

    Article  PubMed  CAS  Google Scholar 

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© 2005 Humana Press Inc., Totowa, NJ

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Cukierman, E. (2005). Cell Migration Analyses Within Fibroblast-Derived 3-D Matrices. In: Guan, JL. (eds) Cell Migration. Methods in Molecular Biology™, vol 294. Humana Press. https://doi.org/10.1385/1-59259-860-9:079

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  • DOI: https://doi.org/10.1385/1-59259-860-9:079

  • Publisher Name: Humana Press

  • Print ISBN: 978-1-58829-382-4

  • Online ISBN: 978-1-59259-860-1

  • eBook Packages: Springer Protocols

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