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The Scar-in-a-Jar: In Vitro Fibrosis Model for Anti-Fibrotic Drug Testing

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Myofibroblasts

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

Abstract

Excessive deposition of type I collagen follows in the wake of chronic inflammation processes in dysregulated tissue healing and causes fibrosis that can ultimately lead to organ failure. While the development of antifibrotic drugs is targeting various upstream events in collagen matrix formation (synthesis, secretion, deposition, stabilization, remodeling), the evaluation of drug effects would use as net read-out of the above effects the presence of a deposited collagen matrix by activated cells, mainly myofibroblasts. Conventional methods comprise lengthy and labor-intensive protocols for the quantification of deposited collagen, some with sensitivity and/or specificity issues. Here we describe the Scar-in-a-Jar assay, an in vitro fibrosis model for anti-fibrotic drug testing that benefits from a substantially accelerated extracellular matrix deposition employing macromolecular crowding and a collagen-producing cell type of choice (e.g., lung fibroblasts like WI-38). The system can be aided by activating compounds such as transforming growth factor-β1, a classical inducer of the myofibroblast phenotype in fibroblasts. Direct image analysis of the well plate not only eliminates the need for matrix extraction or solubilization methods, but also allows for direct imaging and monitoring of phenotypical markers and offers the option for high-content screening applications when adapted to well formats compatible with a screening format.

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Authors and Affiliations

  1. Center for Cell Biology and Tissue Engineering, Institute of Chemistry and Biotechnology, Zurich University of Applied Sciences (ZHAW), Wädenswil, Switzerland

    Simon Stebler & Michael Raghunath

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  1. Simon Stebler
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  2. Michael Raghunath
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Correspondence to Michael Raghunath .

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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Stebler, S., Raghunath, M. (2021). The Scar-in-a-Jar: In Vitro Fibrosis Model for Anti-Fibrotic Drug Testing. 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_11

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  • DOI: https://doi.org/10.1007/978-1-0716-1382-5_11

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-1381-8

  • Online ISBN: 978-1-0716-1382-5

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