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-...
Autores principales: | , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
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MyJove Corporation
2011
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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 |
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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 |