Nothing Special   »   [go: up one dir, main page]

Skip to main content

Double-Hydrogel Substrate as a Model System for Three-Dimensional Cell Culture

  • Protocol
Adhesion Protein Protocols

Part of the book series: Methods in Molecular Biology™ ((MIMB,volume 370))

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.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Subscribe and save

Springer+ Basic
$34.99 /Month
  • Get 10 units per month
  • Download Article/Chapter or eBook
  • 1 Unit = 1 Article or 1 Chapter
  • Cancel anytime
Subscribe now

Buy Now

Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 84.99
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 109.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 109.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

Similar content being viewed by others

References

  1. 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.

    Article  CAS  PubMed  Google Scholar 

  2. 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.

    CAS  Google Scholar 

  3. 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.

    Article  CAS  PubMed  Google Scholar 

  4. 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  CAS  PubMed  Google Scholar 

  5. Grinnell, F. (2003) Fibroblast biology in three-dimensional collagen matrices. Trends Cell Biol. 13, 264–269.

    Article  CAS  PubMed  Google Scholar 

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

    Article  CAS  PubMed  Google Scholar 

  7. Friedl, P. and Brocker, E. B. (2000) The biology of cell locomotion within three-dimensional extracellular matrix. Cell. Mol. Life Sci. 57, 41–64.

    Article  CAS  PubMed  Google Scholar 

  8. 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.

    Article  CAS  PubMed  Google Scholar 

  9. 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.

    Article  CAS  PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Editor information

Editors and Affiliations

Rights and permissions

Reprints 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

Publish with us

Policies and ethics