Fluorescence-activated cell sorting of larval zebrafish muscle stem/progenitor cells following skeletal muscle injury

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Abstract

This chapter describes a protocol for the isolation of larval zebrafish muscle stem/progenitor cells by fluorescence-activated cell sorting (FACS). This method has been successfully applied to isolate pax3a expressing cells 3 days following needle stab skeletal muscle injury. The cell sorting strategy described here can easily be adapted to any cell type at embryonic or larval stages. RNA extracted from the sorted cells can be used for subsequent downstream applications such as quantitative PCR (qPCR), microarrays, or next generation sequencing.

Original languageEnglish
Title of host publicationMyogenesis
Subtitle of host publicationMethods and Protocols
EditorsSissel Beate Ronning
Place of PublicationNew York, NY
PublisherHumana Press
Chapter14
Pages245-254
Number of pages10
ISBN (Electronic)9781493988976
ISBN (Print)9781493988969
DOIs
Publication statusPublished - 2019

Publication series

NameMethods in Molecular Biology
Volume1889
ISSN (Print)1064-3745

Keywords

  • FACS
  • Muscle injury
  • Muscle progenitors
  • Muscle stem cells
  • Needle stab injury
  • Regeneration
  • Satellite cells
  • Skeletal muscle
  • Zebrafish larvae

Cite this

Ratnayake, D., & Currie, P. D. (2019). Fluorescence-activated cell sorting of larval zebrafish muscle stem/progenitor cells following skeletal muscle injury. In S. B. Ronning (Ed.), Myogenesis: Methods and Protocols (pp. 245-254). (Methods in Molecular Biology; Vol. 1889). Humana Press. https://doi.org/10.1007/978-1-4939-8897-6_14