title: Methods for Observing and Quantifying Muscle Satellite Cell Motility and Invasion In Vitro.
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.
authors.affiliation: Division of Biological Sciences and Christopher S. Bond Life Sciences Center, University of Missouri, 1201 Rollins Street, Columbia, MO, 65211 7310, USA.|Developmental Biology Program, Sloan Kettering Institute, New York, NY, USA.
authors.forename: Dane K
authors.initials: DK
authors.lastname: Lund
authors.affiliation: Division of Biological Sciences and Christopher S. Bond Life Sciences Center, University of Missouri, 1201 Rollins Street, Columbia, MO, 65211 7310, USA.|The Kidney Institute, University of Kansas Medical Center, Kansas City, KS, USA.
authors.forename: Patrick
authors.initials: P
authors.lastname: McAnulty
authors.affiliation: Division of Biological Sciences and Christopher S. Bond Life Sciences Center, University of Missouri, 1201 Rollins Street, Columbia, MO, 65211 7310, USA.|Elemental Enzymes, St. Louis, MO, USA.
authors.forename: Ashley L
authors.initials: AL
authors.lastname: Siegel
authors.affiliation: Division of Biological Sciences and Christopher S. Bond Life Sciences Center, University of Missouri, 1201 Rollins Street, Columbia, MO, 65211 7310, USA. cornelisond@missouri.edu.
authors.forename: Ddw
authors.initials: D
authors.lastname: Cornelison
mesh_terms: D002465:Cell Movement; D058948:Cell Tracking / Q000379:methods; D002478:Cells, Cultured; D006801:Humans; D007091:Image Processing, Computer-Assisted; D018485:Muscle Fibers, Skeletal / Q000166:cytology / Q000502:physiology; D018482:Muscle, Skeletal / Q000166:cytology*; D032496:Satellite Cells, Skeletal Muscle / Q000166:cytology* / Q000502:physiology*; D059008:Time-Lapse Imaging / Q000379:methods*
keywords: Cell invasion; Cell motility; Muscle satellite cells; Myofiber culture