Purification of Progenitors from Skeletal Muscle

Skeletal muscle contains multiple progenitor populations of distinct embryonic origins and developmental potential. Myogenic progenitors, usually residing in a "satellite cell position" between the myofiber plasma membrane and the laminin-rich basement membrane that ensheaths it, are self-...

Descripción completa

Detalles Bibliográficos
Autores principales: Yi, Lin, Rossi, Fabio
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MyJove Corporation 2011
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3197294/
https://www.ncbi.nlm.nih.gov/pubmed/21445045
http://dx.doi.org/10.3791/2476
_version_ 1782214295083286528
author Yi, Lin
Rossi, Fabio
author_facet Yi, Lin
Rossi, Fabio
author_sort Yi, Lin
collection PubMed
description Skeletal muscle contains multiple progenitor populations of distinct embryonic origins and developmental potential. Myogenic progenitors, usually residing in a "satellite cell position" between the myofiber plasma membrane and the laminin-rich basement membrane that ensheaths it, are self-renewing cells that are solely committed to the myogenic lineage(1,2). We have recently described a second class of vessel associated progenitors that can generate myofibroblasts and white adipocytes, which responds to damage by efficiently entering proliferation and provides trophic support to myogenic cells during tissue regeneration(3,4). One of the most trusted assays to determine the developmental and regenerative potential of a given cell population relies on their isolation and transplantation(5-7). To this end we have optimized protocols for their purification by flow cytometry from enzymatically dissociated muscle, which we will outline in this article. The populations obtained using this method will contain either myogenic or fibro/adipogenic colony forming cells: no other cell types are capable of expanding in vitro or surviving in vivo delivery. However, when these populations are used immediately after the sort for molecular analysis (e.g qRT-PCR) one must keep in mind that the freshly sorted subsets may contain other contaminant cells that lack the ability of forming colonies or engrafting recipients.
format Online
Article
Text
id pubmed-3197294
institution National Center for Biotechnology Information
language English
publishDate 2011
publisher MyJove Corporation
record_format MEDLINE/PubMed
spelling pubmed-31972942011-10-24 Purification of Progenitors from Skeletal Muscle Yi, Lin Rossi, Fabio J Vis Exp Cellular Biology Skeletal muscle contains multiple progenitor populations of distinct embryonic origins and developmental potential. Myogenic progenitors, usually residing in a "satellite cell position" between the myofiber plasma membrane and the laminin-rich basement membrane that ensheaths it, are self-renewing cells that are solely committed to the myogenic lineage(1,2). We have recently described a second class of vessel associated progenitors that can generate myofibroblasts and white adipocytes, which responds to damage by efficiently entering proliferation and provides trophic support to myogenic cells during tissue regeneration(3,4). One of the most trusted assays to determine the developmental and regenerative potential of a given cell population relies on their isolation and transplantation(5-7). To this end we have optimized protocols for their purification by flow cytometry from enzymatically dissociated muscle, which we will outline in this article. The populations obtained using this method will contain either myogenic or fibro/adipogenic colony forming cells: no other cell types are capable of expanding in vitro or surviving in vivo delivery. However, when these populations are used immediately after the sort for molecular analysis (e.g qRT-PCR) one must keep in mind that the freshly sorted subsets may contain other contaminant cells that lack the ability of forming colonies or engrafting recipients. MyJove Corporation 2011-03-16 /pmc/articles/PMC3197294/ /pubmed/21445045 http://dx.doi.org/10.3791/2476 Text en Copyright © 2011, Journal of Visualized Experiments http://creativecommons.org/licenses/by-nc-nd/3.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution-NonCommercial-NoDerivs 3.0 Unported License. To view a copy of this license, visithttp://creativecommons.org/licenses/by-nc-nd/3.0/
spellingShingle Cellular Biology
Yi, Lin
Rossi, Fabio
Purification of Progenitors from Skeletal Muscle
title Purification of Progenitors from Skeletal Muscle
title_full Purification of Progenitors from Skeletal Muscle
title_fullStr Purification of Progenitors from Skeletal Muscle
title_full_unstemmed Purification of Progenitors from Skeletal Muscle
title_short Purification of Progenitors from Skeletal Muscle
title_sort purification of progenitors from skeletal muscle
topic Cellular Biology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3197294/
https://www.ncbi.nlm.nih.gov/pubmed/21445045
http://dx.doi.org/10.3791/2476
work_keys_str_mv AT yilin purificationofprogenitorsfromskeletalmuscle
AT rossifabio purificationofprogenitorsfromskeletalmuscle