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Quantitative Analysis of Myofibroblast Contraction by Traction Force Microscopy

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Myofibroblasts

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2299))

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Abstract

Myofibroblasts play important roles in physiological processes such as wound healing and tissue repair. While high contractile forces generated by the actomyosin network enable myofibroblasts to physically contract the wound and bring together injured tissue, prolonged and elevated levels of contraction also drive the progression of fibrosis and cancer. However, quantitative mapping of these forces has been difficult due to their extremely low magnitude ranging from 100 pN/μm2 to 2 nN/μm2. Here, we provide a protocol to measure cellular forces exerted on two-dimensional compliant elastic hydrogels. We describe the fabrication of polyacrylamide hydrogels labeled with fluorescent fiducial markers, functionalization of substrates with ECM proteins, setting up the experiment, and imaging procedures. We demonstrate the application of this technique for quantitative analysis of traction forces exerted by myofibroblasts.

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Acknowledgments

Shuying Yang and Fernando R. Valencia contributed equally to this work.

Author information

Author notes

  1. Shuying Yang and Fernando R. Valencia contributed equally to this work.

Authors and Affiliations

  1. Department of Cell and Systems Biology, University of Toronto, Toronto, ON, Canada

    Shuying Yang, Fernando R. Valencia & Sergey V. Plotnikov

  2. Theoretical Physics of Living Matter, Institute of Biological Information Processing and Institute of Advanced Simulation, Forschungszentrum Juelich, Juelich, Germany

    Benedikt Sabass

  3. Department of Veterinary Sciences, Institute for Infectious Diseases and Zoonoses, Ludwig-Maximilians-Universität München, Munich, Germany

    Benedikt Sabass

Authors
  1. Shuying Yang
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  2. Fernando R. Valencia
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  3. Benedikt Sabass
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  4. Sergey V. Plotnikov
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Corresponding author

Correspondence to Sergey V. Plotnikov .

Editor information

Editors and Affiliations

  1. Laboratory of Tissue Repair and Regeneration, Faculty of Dentistry, University of Toronto, Toronto, ON, Canada

    Boris Hinz

  2. Matrix and Mechanobiology Program, Massachusetts General Hospital, Charlestown, MA, USA

    David Lagares

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Yang, S., Valencia, F.R., Sabass, B., Plotnikov, S.V. (2021). Quantitative Analysis of Myofibroblast Contraction by Traction Force Microscopy. In: Hinz, B., Lagares, D. (eds) Myofibroblasts. Methods in Molecular Biology, vol 2299. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1382-5_14

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  • DOI: https://doi.org/10.1007/978-1-0716-1382-5_14

  • Published:

  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-1381-8

  • Online ISBN: 978-1-0716-1382-5

  • eBook Packages: Springer Protocols

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