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Muscle Stem Cells

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Cover of 'Muscle Stem Cells'

Table of Contents

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    Book Overview
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    Chapter 1 Muscle Stem Cells: A Model System for Adult Stem Cell Biology
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    Chapter 2 Isolation of Muscle Stem Cells from Mouse Skeletal Muscle
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    Chapter 3 Primary Mouse Myoblast Purification using Magnetic Cell Separation
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    Chapter 4 Isolation, Culture, and Immunostaining of Skeletal Muscle Myofibers from Wildtype and Nestin-GFP Mice as a Means to Analyze Satellite Cell
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    Chapter 5 Characterization of Drosophila Muscle Stem Cell-Like Adult Muscle Precursors
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    Chapter 6 Using Transgenic Zebrafish to Study Muscle Stem/Progenitor Cells
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    Chapter 7 Muscle Interstitial Cells: A Brief Field Guide to Non-satellite Cell Populations in Skeletal Muscle
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    Chapter 8 Isolation and Characterization of Vessel-Associated Stem/Progenitor Cells from Skeletal Muscle
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    Chapter 9 Fibro/Adipogenic Progenitors (FAPs): Isolation by FACS and Culture
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    Chapter 10 Single Cell Gene Expression Profiling of Skeletal Muscle-Derived Cells
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    Chapter 11 Engraftment of FACS Isolated Muscle Stem Cells into Injured Skeletal Muscle
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    Chapter 12 Transplantation of Skeletal Muscle Stem Cells
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    Chapter 13 Simultaneous Measurement of Mitochondrial and Glycolytic Activity in Quiescent Muscle Stem Cells
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    Chapter 14 Monitoring Autophagy in Muscle Stem Cells
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    Chapter 15 Mimicking Muscle Stem Cell Quiescence in Culture: Methods for Synchronization in Reversible Arrest
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    Chapter 16 Methods for Observing and Quantifying Muscle Satellite Cell Motility and Invasion In Vitro
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    Chapter 17 Effects of Macrophage Conditioned-Medium on Murine and Human Muscle Cells: Analysis of Proliferation, Differentiation, and Fusion
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    Chapter 18 Optimization of Satellite Cell Culture Through Biomaterials
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    Chapter 19 Systematic Identification of Genes Regulating Muscle Stem Cell Self-Renewal and Differentiation
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    Chapter 20 Bioinformatics for Novel Long Intergenic Noncoding RNA (lincRNA) Identification in Skeletal Muscle Cells
Attention for Chapter 4: Isolation, Culture, and Immunostaining of Skeletal Muscle Myofibers from Wildtype and Nestin-GFP Mice as a Means to Analyze Satellite Cell
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Chapter title
Isolation, Culture, and Immunostaining of Skeletal Muscle Myofibers from Wildtype and Nestin-GFP Mice as a Means to Analyze Satellite Cell
Chapter number 4
Book title
Muscle Stem Cells
Published in
Methods in molecular biology, March 2017
DOI 10.1007/978-1-4939-6771-1_4
Pubmed ID
Book ISBNs
978-1-4939-6769-8, 978-1-4939-6771-1
Authors

Pascal Stuelsatz, Paul Keire, Zipora Yablonka-Reuveni, Stuelsatz, Pascal, Keire, Paul, Yablonka-Reuveni, Zipora

Editors

Eusebio Perdiguero, DDW Cornelison

Abstract

Multinucleated myofibers, the functional contractile units of adult skeletal muscle, harbor mononuclear Pax7(+) myogenic progenitors on their surface between the myofiber basal lamina and plasmalemma. These progenitors, known as satellite cells, are the primary myogenic stem cells in adult muscle. This chapter describes our laboratory protocols for isolating, culturing, and immunostaining intact myofibers from mouse skeletal muscle as a means for studying satellite cell dynamics. The first protocol discusses myofiber isolation from the flexor digitorum brevis (FDB) muscle. These short myofibers are plated in dishes coated with PureCol collagen (formerly known as Vitrogen) and maintained in a mitogen-poor medium (± supplemental growth factors). Employing such conditions, satellite cells remain at the surface of the parent myofiber while synchronously undergoing a limited number of proliferative cycles and rapidly differentiate. The second protocol discusses the isolation of longer myofibers from the extensor digitorum longus (EDL) muscle. These EDL myofibers are routinely plated individually as adherent myofibers in wells coated with Matrigel and maintained in a mitogen-rich medium, conditions in which satellite cells migrate away from the parent myofiber, proliferate extensively, and generate numerous differentiating progeny. Alternatively, these EDL myofibers can be plated as non-adherent myofibers in uncoated wells and maintained in a mitogen-poor medium (± supplemental growth factors), conditions that retain satellite cell progeny at the myofiber niche similar to the FDB myofiber cultures. However, the adherent myofiber format is our preferred choice for monitoring satellite cells in freshly isolated (Time 0) myofibers. We conclude this chapter by promoting the Nestin-GFP transgenic mouse as an efficient tool for direct analysis of satellite cells in isolated myofibers. While satellite cells have been often detected by their expression of the Pax7 protein or the Myf5(nLacZ) knockin reporter (approaches that are also detailed herein), the Nestin-GFP reporter distinctively permits quantification of satellite cells in live myofibers, which enables linking initial Time 0 numbers and subsequent performance upon culturing. We additionally point out to the implementation of the Nestin-GFP transgene for monitoring other selective cell lineages as illustrated by GFP expression in capillaries, endothelial tubes and neuronal cells. Myofibers from other types of muscles, such as diaphragm, masseter, and extraocular, can also be isolated and analyzed using protocols described herein. Collectively, this chapter provides essential tools for studying satellite cells in their native position and their interplay with the parent myofiber.

Twitter Demographics

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Mendeley readers

The data shown below were compiled from readership statistics for 27 Mendeley readers of this research output. Click here to see the associated Mendeley record.

Geographical breakdown

Country Count As %
Unknown 27 100%

Demographic breakdown

Readers by professional status Count As %
Student > Ph. D. Student 6 22%
Student > Master 5 19%
Researcher 5 19%
Student > Doctoral Student 3 11%
Student > Bachelor 2 7%
Other 3 11%
Unknown 3 11%
Readers by discipline Count As %
Biochemistry, Genetics and Molecular Biology 12 44%
Agricultural and Biological Sciences 4 15%
Medicine and Dentistry 3 11%
Neuroscience 2 7%
Sports and Recreations 1 4%
Other 1 4%
Unknown 4 15%

Attention Score in Context

This research output has an Altmetric Attention Score of 1. This is our high-level measure of the quality and quantity of online attention that it has received. This Attention Score, as well as the ranking and number of research outputs shown below, was calculated when the research output was last mentioned on 11 March 2017.
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#7,957,187
of 9,178,972 outputs
Outputs from Methods in molecular biology
#4,872
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Outputs of similar age
#215,092
of 254,195 outputs
Outputs of similar age from Methods in molecular biology
#8
of 23 outputs
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