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A novel approach for the isolation and long-term expansion of pure satellite cells based on ice-cold treatment
Satellite cells (SCs) are muscle stem cells capable of regenerating injured muscle. The study of their functional potential depends on the availability of methods for the isolation and expansion of pure SCs with preserved myogenic properties after serial passages in vitro. Here, we describe the ice-...
Autores principales: | , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
BioMed Central
2021
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7968259/ https://www.ncbi.nlm.nih.gov/pubmed/33731194 http://dx.doi.org/10.1186/s13395-021-00261-w |
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author | Benedetti, Anna Cera, Gianluca De Meo, Daniele Villani, Ciro Bouche, Marina Lozanoska-Ochser, Biliana |
author_facet | Benedetti, Anna Cera, Gianluca De Meo, Daniele Villani, Ciro Bouche, Marina Lozanoska-Ochser, Biliana |
author_sort | Benedetti, Anna |
collection | PubMed |
description | Satellite cells (SCs) are muscle stem cells capable of regenerating injured muscle. The study of their functional potential depends on the availability of methods for the isolation and expansion of pure SCs with preserved myogenic properties after serial passages in vitro. Here, we describe the ice-cold treatment (ICT) method, which is a simple, economical, and efficient method for the isolation and in vitro expansion of highly pure mouse and human SCs. It involves a brief (15–30 min) incubation on ice (0 °C) of a dish containing a heterogeneous mix of adherent muscle mononuclear cells, which leads to the detachment of only the SCs, and gives rise to cultures of superior purity compared to other commonly used isolation methods. The ICT method doubles up as a gentle passaging technique, allowing SC expansion over extended periods of time without compromising their proliferation and differentiation potential. Moreover, SCs isolated and expanded using the ICT method are capable of regenerating injured muscle in vivo. The ICT method involves minimal cell manipulation, does not require any expertise or expensive reagents, it is fast, and highly reproducible, and greatly reduces the number of animals or human biopsies required in order to obtain sufficient number of SCs. The cost-effectiveness, accessibility, and technical simplicity of this method, as well as its remarkable efficiency, will no doubt accelerate SC basic and translational research bringing their therapeutic use closer to the clinic. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s13395-021-00261-w. |
format | Online Article Text |
id | pubmed-7968259 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-79682592021-03-22 A novel approach for the isolation and long-term expansion of pure satellite cells based on ice-cold treatment Benedetti, Anna Cera, Gianluca De Meo, Daniele Villani, Ciro Bouche, Marina Lozanoska-Ochser, Biliana Skelet Muscle Methodology Satellite cells (SCs) are muscle stem cells capable of regenerating injured muscle. The study of their functional potential depends on the availability of methods for the isolation and expansion of pure SCs with preserved myogenic properties after serial passages in vitro. Here, we describe the ice-cold treatment (ICT) method, which is a simple, economical, and efficient method for the isolation and in vitro expansion of highly pure mouse and human SCs. It involves a brief (15–30 min) incubation on ice (0 °C) of a dish containing a heterogeneous mix of adherent muscle mononuclear cells, which leads to the detachment of only the SCs, and gives rise to cultures of superior purity compared to other commonly used isolation methods. The ICT method doubles up as a gentle passaging technique, allowing SC expansion over extended periods of time without compromising their proliferation and differentiation potential. Moreover, SCs isolated and expanded using the ICT method are capable of regenerating injured muscle in vivo. The ICT method involves minimal cell manipulation, does not require any expertise or expensive reagents, it is fast, and highly reproducible, and greatly reduces the number of animals or human biopsies required in order to obtain sufficient number of SCs. The cost-effectiveness, accessibility, and technical simplicity of this method, as well as its remarkable efficiency, will no doubt accelerate SC basic and translational research bringing their therapeutic use closer to the clinic. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s13395-021-00261-w. BioMed Central 2021-03-17 /pmc/articles/PMC7968259/ /pubmed/33731194 http://dx.doi.org/10.1186/s13395-021-00261-w Text en © The Author(s) 2021 Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data. |
spellingShingle | Methodology Benedetti, Anna Cera, Gianluca De Meo, Daniele Villani, Ciro Bouche, Marina Lozanoska-Ochser, Biliana A novel approach for the isolation and long-term expansion of pure satellite cells based on ice-cold treatment |
title | A novel approach for the isolation and long-term expansion of pure satellite cells based on ice-cold treatment |
title_full | A novel approach for the isolation and long-term expansion of pure satellite cells based on ice-cold treatment |
title_fullStr | A novel approach for the isolation and long-term expansion of pure satellite cells based on ice-cold treatment |
title_full_unstemmed | A novel approach for the isolation and long-term expansion of pure satellite cells based on ice-cold treatment |
title_short | A novel approach for the isolation and long-term expansion of pure satellite cells based on ice-cold treatment |
title_sort | novel approach for the isolation and long-term expansion of pure satellite cells based on ice-cold treatment |
topic | Methodology |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7968259/ https://www.ncbi.nlm.nih.gov/pubmed/33731194 http://dx.doi.org/10.1186/s13395-021-00261-w |
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