Abstract
When cultured on two-dimensional surfaces, most adherent cells show profound differences from those in their native habitats. In addition to chemical factors, it is likely that both physical parameters, such as substrate rigidity, and topographical factors, such as the asymmetry in integrin anchorage, play a major role in the differences. We have designed a simple culture system that provides flexible, adhesive substrates for both dorsal and ventral cell surfaces. Fibroblasts in this system show the spindle or stellate morphology found in native tissues. The ease of preparation, versatility, and optical quality of this model system should greatly facilitate the understanding of cellular behavior and functions in vivo.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Similar content being viewed by others
References
Meier, S. and Hay, E. D. (1975) Stimulation of corneal differentiation by interaction between cell surface and extracellular matrix. I. Morphometric analysis of transfilter “induction.” J. Cell Biol. 66, 275–291.
Mauchamp, J., Chambard, M., Verrier, B., et al. (1987) Epithelial cell polarization in culture: orientation of cell polarity and expression of specific functions, studied with cultured thyroid cells. J. Cell Sci. 8, 345–358.
Cook, J. R., Crute, B. E., Patrone, L. M., Gabriels, J., Lane, M. E., and Van Buskirk, R. G. (1989) Microporosity of the substratum regulates differentiation of MDCK cells in vitro. In Vitro Cell Dev. Biol. 25, 914–922.
Schmeichel, K. L. and Bissell, M. J. (2003) Modeling tissue-specific signaling and organ function in three dimensions. J. Cell Sci. 116, 2377–2388.
Grinnell, F. (2003) Fibroblast biology in three-dimensional collagen matrices. Trends Cell Biol. 13, 264–269.
Cukierman, E., Pankov, R., Stevens, D. R., and Yamada, K. M. (2001) Taking cellmatrix adhesions to the third dimension. Science 294, 1708–1712.
Friedl, P. and Brocker, E. B. (2000) The biology of cell locomotion within three-dimensional extracellular matrix. Cell. Mol. Life Sci. 57, 41–64.
Beningo, K. A., Dembo, M., and Wang, Y.-L. (2004) Responses of fibroblasts to anchorage of dorsal extracellular matrix receptors. Proc. Natl. Acad. Sci. USA 101, 18,024–18,029.
Beningo, K. A., Lo, C.-M., and Wang, Y.-L. (2002) Flexible polyacrylamide substrata for the analysis of mechanical interactions at cell-substratum adhesions. Methods Cell Biol. 69, 325–339.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2007 Humana Press Inc., Totowa, NJ
About this protocol
Cite this protocol
Beningo, K.A., Wang, Yl. (2007). Double-Hydrogel Substrate as a Model System for Three-Dimensional Cell Culture. In: Coutts, A.S. (eds) Adhesion Protein Protocols. Methods in Molecular Biology™, vol 370. Humana Press. https://doi.org/10.1007/978-1-59745-353-0_14
Download citation
DOI: https://doi.org/10.1007/978-1-59745-353-0_14
Publisher Name: Humana Press
Print ISBN: 978-1-58829-533-0
Online ISBN: 978-1-59745-353-0
eBook Packages: Springer Protocols