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

Research output: Chapter in Book/Report/Conference proceedingChapter (Book)Otherpeer-review

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). New York, NY: Humana Press. https://doi.org/10.1007/978-1-4939-8897-6_14
Ratnayake, Dhanushika ; Currie, Peter D. / Fluorescence-activated cell sorting of larval zebrafish muscle stem/progenitor cells following skeletal muscle injury. Myogenesis: Methods and Protocols. editor / Sissel Beate Ronning. New York, NY : Humana Press, 2019. pp. 245-254 (Methods in Molecular Biology).
@inbook{be82fc7575774717b36e2b122c193364,
title = "Fluorescence-activated cell sorting of larval zebrafish muscle stem/progenitor cells following skeletal muscle injury",
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.",
keywords = "FACS, Muscle injury, Muscle progenitors, Muscle stem cells, Needle stab injury, Regeneration, Satellite cells, Skeletal muscle, Zebrafish larvae",
author = "Dhanushika Ratnayake and Currie, {Peter D.}",
year = "2019",
doi = "10.1007/978-1-4939-8897-6_14",
language = "English",
isbn = "9781493988969",
series = "Methods in Molecular Biology",
publisher = "Humana Press",
pages = "245--254",
editor = "Ronning, {Sissel Beate}",
booktitle = "Myogenesis",
address = "United States of America",

}

Ratnayake, D & Currie, PD 2019, Fluorescence-activated cell sorting of larval zebrafish muscle stem/progenitor cells following skeletal muscle injury. in SB Ronning (ed.), Myogenesis: Methods and Protocols. Methods in Molecular Biology, vol. 1889, Humana Press, New York, NY, pp. 245-254. https://doi.org/10.1007/978-1-4939-8897-6_14

Fluorescence-activated cell sorting of larval zebrafish muscle stem/progenitor cells following skeletal muscle injury. / Ratnayake, Dhanushika; Currie, Peter D.

Myogenesis: Methods and Protocols. ed. / Sissel Beate Ronning. New York, NY : Humana Press, 2019. p. 245-254 (Methods in Molecular Biology; Vol. 1889).

Research output: Chapter in Book/Report/Conference proceedingChapter (Book)Otherpeer-review

TY - CHAP

T1 - Fluorescence-activated cell sorting of larval zebrafish muscle stem/progenitor cells following skeletal muscle injury

AU - Ratnayake, Dhanushika

AU - Currie, Peter D.

PY - 2019

Y1 - 2019

N2 - 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.

AB - 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.

KW - FACS

KW - Muscle injury

KW - Muscle progenitors

KW - Muscle stem cells

KW - Needle stab injury

KW - Regeneration

KW - Satellite cells

KW - Skeletal muscle

KW - Zebrafish larvae

UR - http://www.scopus.com/inward/record.url?scp=85055613383&partnerID=8YFLogxK

U2 - 10.1007/978-1-4939-8897-6_14

DO - 10.1007/978-1-4939-8897-6_14

M3 - Chapter (Book)

SN - 9781493988969

T3 - Methods in Molecular Biology

SP - 245

EP - 254

BT - Myogenesis

A2 - Ronning, Sissel Beate

PB - Humana Press

CY - New York, NY

ER -

Ratnayake D, Currie PD. Fluorescence-activated cell sorting of larval zebrafish muscle stem/progenitor cells following skeletal muscle injury. In Ronning SB, editor, Myogenesis: Methods and Protocols. New York, NY: Humana Press. 2019. p. 245-254. (Methods in Molecular Biology). https://doi.org/10.1007/978-1-4939-8897-6_14