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Methods for Observing and Quantifying Muscle Satellite Cell Motility and Invasion In Vitro

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Muscle Stem Cells

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1556))

Abstract

Motility and/or chemotaxis of satellite cells has been suggested or observed in multiple in vitro and in vivo contexts. Satellite cell motility also affects the efficiency of muscle regeneration, particularly in the context of engrafted exogenous cells. Consequently, there is keen interest in determining what cell-autonomous and environmental factors influence satellite cell motility and chemotaxis in vitro and in vivo. In addition, the ability of activated satellite cells to relocate in vivo would suggest that they must be able to invade and transit through the extracellular matrix (ECM), which is supported by studies in which alteration or addition of matrix metalloprotease (MMP) activity enhanced the spread of engrafted satellite cells. However, despite its potential importance, analysis of satellite cell motility or invasion quantitatively even in an in vitro setting can be difficult; one of the most powerful techniques for overcoming these difficulties is timelapse microscopy. Identification and longitudinal evaluation of individual cells over time permits not only quantification of variations in motility due to intrinsic or extrinsic factors, it permits observation and analysis of other (frequently unsuspected) cellular activities as well. We describe here three protocols developed in our group for quantitatively analyzing satellite cell motility over time in two dimensions on purified ECM substrates, in three dimensions on a living myofiber, and in three dimensions through an artificial matrix.

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Change history

  • 20 March 2018

    Correction to: Eusebio Perdiguero and DDW Cornelison (eds.), Muscle Stem Cells: Methods and Protocols, Methods in Molecular Biology, vol. 1556, https://doi.org/10.1007/978-1-4939-6771-1

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Author notes

    Authors and Affiliations

    1. Division of Biological Sciences and Christopher S. Bond Life Sciences Center, University of Missouri, 1201 Rollins Street, Columbia, MO, 65211 7310, USA

      Dane K. Lund, Patrick McAnulty, Ashley L. Siegel & DDW Cornelison

    2. Developmental Biology Program, Sloan Kettering Institute, New York, NY, USA

      Dane K. Lund

    3. The Kidney Institute, University of Kansas Medical Center, Kansas City, KS, USA

      Patrick McAnulty

    4. Elemental Enzymes, St. Louis, MO, USA

      Ashley L. Siegel

    Authors
    1. Dane K. Lund
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    2. Patrick McAnulty
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    3. Ashley L. Siegel
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    4. DDW Cornelison
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    Corresponding author

    Correspondence to DDW Cornelison .

    Editor information

    Editors and Affiliations

    1. Cell Biology Group, Department of Experimental and Health Sciences, University Pompeu Fabra (UPF), Barcelona, Spain

      Eusebio Perdiguero

    2. Division of Biological Sciences and Christopher S. Bond Life Sciences Center, University of Missouri, Columbia, MO, USA

      DDW Cornelison

    1 Electronic Supplementary Material

    Below is the link to the electronic supplementary material.

    A single-plane timelapse recording through a collagen I matrix showing invasion of human satellite cells over 24 h (M4V 5732 KB)

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    Lund, D.K., McAnulty, P., Siegel, A.L., Cornelison, D. (2017). Methods for Observing and Quantifying Muscle Satellite Cell Motility and Invasion In Vitro. In: Perdiguero, E., Cornelison, D. (eds) Muscle Stem Cells. Methods in Molecular Biology, vol 1556. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-6771-1_16

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    • DOI: https://doi.org/10.1007/978-1-4939-6771-1_16

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    • Publisher Name: Humana, New York, NY

    • Print ISBN: 978-1-4939-6769-8

    • Online ISBN: 978-1-4939-6771-1

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