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Myofibroblast Markers and Microscopy Detection Methods in Cell Culture and Histology

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Myofibroblasts

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

Abstract

The identification of myofibroblasts is essential for mechanistic in vitro studies, cell-based drug tests, and to assess the level of fibrosis in experimental animal or human fibrosis. The name myo-fibroblast was chosen in 1971 to express that the formation of contractile features-stress fibers is the essential criterion to define these cells. Additional neo-expression of α-smooth muscle actin (α-SMA) in stress fibers has become the most widely used molecular marker. Here, we briefly introduce the concept of different myofibroblast activation states, of which the highly contractile α-SMA-positive phenotype represents a most advanced functional stage. We provide targeted immunofluorescence protocols to assess this phenotype, and publicly accessible image analysis tools to quantify the level of myofibroblast activation in culture and in tissues.

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Acknowledgments

We would like to thank to Dr. Giulio Gabbiani for kindly providing α-SMA antibody and the staff of the Collaborative Advanced Microscopy Labs of Dentistry (CAMiLoD) (Faculty of Dentistry, University of Toronto) for technical support. Schemes were produced with Biorender (biorender.com). The research of BH is supported by a foundation grant from the Canadian Institutes of Health Research (#375597) and support from the John Evans Leadership funds (#36050 and #38861) and innovation funds (“Fibrosis Network, #36349”) from the Canada Foundation for Innovation (CFI) and the Ontario Research Fund (ORF). FY is recipient of a Mary H. Beatty Fellowship from the School of Graduate Studies, University of Toronto. Fereshteh Younesi and Dong Ok Son contributed equally to this work.

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Author notes

  1. Fereshteh Younesi and Dong Ok Son authors contributed equally to this work.

Authors and Affiliations

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

    Fereshteh S. Younesi, Dong Ok Son, Joao Firmino & Boris Hinz

  2. Collaborative Advanced Microscopy Laboratories of Dentistry (CAMiLoD), Faculty of Dentistry, University of Toronto, Toronto, ON, Canada

    Joao Firmino & Boris Hinz

Authors
  1. Fereshteh S. Younesi
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  2. Dong Ok Son
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  3. Joao Firmino
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  4. Boris Hinz
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Correspondence to Boris Hinz .

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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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Younesi, F.S., Son, D.O., Firmino, J., Hinz, B. (2021). Myofibroblast Markers and Microscopy Detection Methods in Cell Culture and Histology. 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_3

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  • DOI: https://doi.org/10.1007/978-1-0716-1382-5_3

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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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