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A Three-Dimensional Culture Model of Reversibly Quiescent Myogenic Cells

Satellite cells (SC) are the stem cells of skeletal muscles. They are quiescent in adult animals but resume proliferation to allow muscle hypertrophy or regeneration after injury. The mechanisms balancing quiescence, self-renewal, and differentiation of SC are difficult to analyze in vivo owing to t...

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Autores principales: Aguanno, Salvatore, Petrelli, Claudia, Di Siena, Sara, De Angelis, Luciana, Pellegrini, Manuela, Naro, Fabio
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Hindawi 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6885280/
https://www.ncbi.nlm.nih.gov/pubmed/31827532
http://dx.doi.org/10.1155/2019/7548160
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author Aguanno, Salvatore
Petrelli, Claudia
Di Siena, Sara
De Angelis, Luciana
Pellegrini, Manuela
Naro, Fabio
author_facet Aguanno, Salvatore
Petrelli, Claudia
Di Siena, Sara
De Angelis, Luciana
Pellegrini, Manuela
Naro, Fabio
author_sort Aguanno, Salvatore
collection PubMed
description Satellite cells (SC) are the stem cells of skeletal muscles. They are quiescent in adult animals but resume proliferation to allow muscle hypertrophy or regeneration after injury. The mechanisms balancing quiescence, self-renewal, and differentiation of SC are difficult to analyze in vivo owing to their complexity and in vitro because the staminal character of SC is lost when they are removed from the niche and is not adequately reproduced in the culture models currently available. To overcome these difficulties, we set up a culture model of the myogenic C2C12 cell line in suspension. When C2C12 cells are cultured in suspension, they enter a state of quiescence and form three-dimensional aggregates (myospheres) that produce the extracellular matrix and express markers of quiescent SC. In the initial phase of culture, a portion of the cells fuses in syncytia and abandons the myospheres. The remaining cells are mononucleated and quiescent but resume proliferation and differentiation when plated in a monolayer. The notch pathway controls the quiescent state of the cells as shown by the fact that its inhibition leads to the resumption of differentiation. Within this context, notch3 appears to play a central role in the activity of this pathway since the expression of notch1 declines soon after aggregation. In summary, the culture model of C2C12 in suspension may be used to study the cellular interactions of muscle stem cells and the pathways controlling SC quiescence entrance and maintenance.
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spelling pubmed-68852802019-12-11 A Three-Dimensional Culture Model of Reversibly Quiescent Myogenic Cells Aguanno, Salvatore Petrelli, Claudia Di Siena, Sara De Angelis, Luciana Pellegrini, Manuela Naro, Fabio Stem Cells Int Research Article Satellite cells (SC) are the stem cells of skeletal muscles. They are quiescent in adult animals but resume proliferation to allow muscle hypertrophy or regeneration after injury. The mechanisms balancing quiescence, self-renewal, and differentiation of SC are difficult to analyze in vivo owing to their complexity and in vitro because the staminal character of SC is lost when they are removed from the niche and is not adequately reproduced in the culture models currently available. To overcome these difficulties, we set up a culture model of the myogenic C2C12 cell line in suspension. When C2C12 cells are cultured in suspension, they enter a state of quiescence and form three-dimensional aggregates (myospheres) that produce the extracellular matrix and express markers of quiescent SC. In the initial phase of culture, a portion of the cells fuses in syncytia and abandons the myospheres. The remaining cells are mononucleated and quiescent but resume proliferation and differentiation when plated in a monolayer. The notch pathway controls the quiescent state of the cells as shown by the fact that its inhibition leads to the resumption of differentiation. Within this context, notch3 appears to play a central role in the activity of this pathway since the expression of notch1 declines soon after aggregation. In summary, the culture model of C2C12 in suspension may be used to study the cellular interactions of muscle stem cells and the pathways controlling SC quiescence entrance and maintenance. Hindawi 2019-11-11 /pmc/articles/PMC6885280/ /pubmed/31827532 http://dx.doi.org/10.1155/2019/7548160 Text en Copyright © 2019 Salvatore Aguanno et al. http://creativecommons.org/licenses/by/4.0/ This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Research Article
Aguanno, Salvatore
Petrelli, Claudia
Di Siena, Sara
De Angelis, Luciana
Pellegrini, Manuela
Naro, Fabio
A Three-Dimensional Culture Model of Reversibly Quiescent Myogenic Cells
title A Three-Dimensional Culture Model of Reversibly Quiescent Myogenic Cells
title_full A Three-Dimensional Culture Model of Reversibly Quiescent Myogenic Cells
title_fullStr A Three-Dimensional Culture Model of Reversibly Quiescent Myogenic Cells
title_full_unstemmed A Three-Dimensional Culture Model of Reversibly Quiescent Myogenic Cells
title_short A Three-Dimensional Culture Model of Reversibly Quiescent Myogenic Cells
title_sort three-dimensional culture model of reversibly quiescent myogenic cells
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6885280/
https://www.ncbi.nlm.nih.gov/pubmed/31827532
http://dx.doi.org/10.1155/2019/7548160
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