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

  • Dane K. Lund
  • Patrick McAnulty
  • Ashley L. Siegel
  • DDW CornelisonEmail author
Protocol
Part of the Methods in Molecular Biology book series (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.

Key words

Muscle satellite cells Cell motility Myofiber culture Cell invasion 

Supplementary material

Supplemental Movie 1

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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Copyright information

© Springer Science+Business Media LLC 2017

Open Access This chapter is distributed under the terms of the Creative Commons Attribution Noncommercial License, which permits any noncommercial use, distribution, and reproduction in any medium, provided the original author(s) and source are credited.

Authors and Affiliations

  • Dane K. Lund
    • 1
    • 2
  • Patrick McAnulty
    • 1
    • 3
  • Ashley L. Siegel
    • 1
    • 4
  • DDW Cornelison
    • 1
    Email author
  1. 1.Division of Biological Sciences and Christopher S. Bond Life Sciences CenterUniversity of MissouriColumbiaUSA
  2. 2.Developmental Biology ProgramSloan Kettering InstituteNew YorkUSA
  3. 3.The Kidney InstituteUniversity of Kansas Medical CenterKansas CityUSA
  4. 4.Elemental EnzymesSt. LouisUSA

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