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Generation of 3D Fibroblast-Derived Extracellular Matrix and Analysis of Tumor Cell-Matrix Interactions and Signaling

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Imaging Cell Signaling

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2800))

Abstract

Fibroblasts are the major producers of the extracellular matrix and regulate its organization. Aberrant signaling in diseases such as fibrosis and cancer can impact the deposition of the matrix proteins, which can in turn act as an adhesion scaffold and signaling reservoir promoting disease progression. To study the composition and organization of the extracellular matrix as well as its interactions with (tumor) cells, this protocol describes the generation and analysis of 3D fibroblast-derived matrices and the investigation of (tumor) cells seeded onto the 3D scaffolds by immunofluorescent imaging and cell adhesion, colony formation, migration, and invasion/transmigration assays.

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Acknowledgments

This work was supported in parts by grant 016/S/20 from the British Skin Foundation awarded to Ute Jungwirth, by grant BB/T012978/1 from the Biotechnology and Biological Sciences Research Council BBSRC/UKRI awarded to Gernot Walko, and by grant MR/W006308/1 for the GW4 BIOMED2 DTP awarded to the Universities of Bath, Bristol, Cardiff and Exeter from the Medical Research Council (MRC)/UKRI. Figure 1 was partly generated using Servier Medical Art, provided by Servier, licensed under a Creative Commons Attribution 3.0 unported license.

Author information

Authors and Affiliations

  1. Department of Life Sciences, University of Bath, Bath, UK

    Emily Lay, Tressan Grant, Gernot Walko & Ute Jungwirth

  2. School of Cellular and Molecular Medicine, University of Bristol, Bristol, UK

    Tressan Grant

  3. Barts Centre for Squamous Cancer, Institute of Dentistry, Queen Mary University of London, London, UK

    Gernot Walko

  4. Centre for Oral Immunobiology and Regenerative Medicine, Institute of Dentistry, Queen Mary University of London, London, UK

    Gernot Walko

  5. Cancer Research Horizons Therapeutic Innovation, Newcastle Drug Discovery Group, Translational and Clinical Research Institute, Newcastle University, Newcastle, UK

    Ute Jungwirth

Authors
  1. Emily Lay
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  2. Tressan Grant
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  3. Gernot Walko
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  4. Ute Jungwirth
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Corresponding author

Correspondence to Ute Jungwirth .

Editor information

Editors and Affiliations

  1. School of Cellular and Molecular Medicine, University of Bristol, Bristol, UK

    Christoph Wuelfing

  2. Computational Biology Department, Carnegie Mellon University, Pittsburgh, PA, USA

    Robert F. Murphy

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© 2024 The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature

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Lay, E., Grant, T., Walko, G., Jungwirth, U. (2024). Generation of 3D Fibroblast-Derived Extracellular Matrix and Analysis of Tumor Cell-Matrix Interactions and Signaling. In: Wuelfing, C., Murphy, R.F. (eds) Imaging Cell Signaling. Methods in Molecular Biology, vol 2800. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3834-7_2

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  • DOI: https://doi.org/10.1007/978-1-0716-3834-7_2

  • Published:

  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-3833-0

  • Online ISBN: 978-1-0716-3834-7

  • eBook Packages: Springer Protocols

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