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Novel integrated workflow allows production and in-depth quality assessment of multifactorial reprogrammed skeletal muscle cells from human stem cells

Skeletal muscle tissue engineering aims at generating biological substitutes that restore, maintain or improve normal muscle function; however, the quality of cells produced by current protocols remains insufficient. Here, we developed a multifactor-based protocol that combines adenovector (AdV)-med...

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Autores principales: Faustino, Dinis, Brinkmeier, Heinrich, Logotheti, Stella, Jonitz-Heincke, Anika, Yilmaz, Hande, Takan, Isil, Peters, Kirsten, Bader, Rainer, Lang, Hermann, Pavlopoulou, Athanasia, Pützer, Brigitte M., Spitschak, Alf
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Springer International Publishing 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8993739/
https://www.ncbi.nlm.nih.gov/pubmed/35396689
http://dx.doi.org/10.1007/s00018-022-04264-8
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author Faustino, Dinis
Brinkmeier, Heinrich
Logotheti, Stella
Jonitz-Heincke, Anika
Yilmaz, Hande
Takan, Isil
Peters, Kirsten
Bader, Rainer
Lang, Hermann
Pavlopoulou, Athanasia
Pützer, Brigitte M.
Spitschak, Alf
author_facet Faustino, Dinis
Brinkmeier, Heinrich
Logotheti, Stella
Jonitz-Heincke, Anika
Yilmaz, Hande
Takan, Isil
Peters, Kirsten
Bader, Rainer
Lang, Hermann
Pavlopoulou, Athanasia
Pützer, Brigitte M.
Spitschak, Alf
author_sort Faustino, Dinis
collection PubMed
description Skeletal muscle tissue engineering aims at generating biological substitutes that restore, maintain or improve normal muscle function; however, the quality of cells produced by current protocols remains insufficient. Here, we developed a multifactor-based protocol that combines adenovector (AdV)-mediated MYOD expression, small molecule inhibitor and growth factor treatment, and electrical pulse stimulation (EPS) to efficiently reprogram different types of human-derived multipotent stem cells into physiologically functional skeletal muscle cells (SMCs). The protocol was complemented through a novel in silico workflow that allows for in-depth estimation and potentially optimization of the quality of generated muscle tissue, based on the transcriptomes of transdifferentiated cells. We additionally patch-clamped phenotypic SMCs to associate their bioelectrical characteristics with their transcriptome reprogramming. Overall, we set up a comprehensive and dynamic approach at the nexus of viral vector-based technology, bioinformatics, and electrophysiology that facilitates production of high-quality skeletal muscle cells and can guide iterative cycles to improve myo-differentiation protocols. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s00018-022-04264-8.
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spelling pubmed-89937392022-04-22 Novel integrated workflow allows production and in-depth quality assessment of multifactorial reprogrammed skeletal muscle cells from human stem cells Faustino, Dinis Brinkmeier, Heinrich Logotheti, Stella Jonitz-Heincke, Anika Yilmaz, Hande Takan, Isil Peters, Kirsten Bader, Rainer Lang, Hermann Pavlopoulou, Athanasia Pützer, Brigitte M. Spitschak, Alf Cell Mol Life Sci Original Article Skeletal muscle tissue engineering aims at generating biological substitutes that restore, maintain or improve normal muscle function; however, the quality of cells produced by current protocols remains insufficient. Here, we developed a multifactor-based protocol that combines adenovector (AdV)-mediated MYOD expression, small molecule inhibitor and growth factor treatment, and electrical pulse stimulation (EPS) to efficiently reprogram different types of human-derived multipotent stem cells into physiologically functional skeletal muscle cells (SMCs). The protocol was complemented through a novel in silico workflow that allows for in-depth estimation and potentially optimization of the quality of generated muscle tissue, based on the transcriptomes of transdifferentiated cells. We additionally patch-clamped phenotypic SMCs to associate their bioelectrical characteristics with their transcriptome reprogramming. Overall, we set up a comprehensive and dynamic approach at the nexus of viral vector-based technology, bioinformatics, and electrophysiology that facilitates production of high-quality skeletal muscle cells and can guide iterative cycles to improve myo-differentiation protocols. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s00018-022-04264-8. Springer International Publishing 2022-04-09 2022 /pmc/articles/PMC8993739/ /pubmed/35396689 http://dx.doi.org/10.1007/s00018-022-04264-8 Text en © The Author(s) 2022 https://creativecommons.org/licenses/by/4.0/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/ (https://creativecommons.org/licenses/by/4.0/) .
spellingShingle Original Article
Faustino, Dinis
Brinkmeier, Heinrich
Logotheti, Stella
Jonitz-Heincke, Anika
Yilmaz, Hande
Takan, Isil
Peters, Kirsten
Bader, Rainer
Lang, Hermann
Pavlopoulou, Athanasia
Pützer, Brigitte M.
Spitschak, Alf
Novel integrated workflow allows production and in-depth quality assessment of multifactorial reprogrammed skeletal muscle cells from human stem cells
title Novel integrated workflow allows production and in-depth quality assessment of multifactorial reprogrammed skeletal muscle cells from human stem cells
title_full Novel integrated workflow allows production and in-depth quality assessment of multifactorial reprogrammed skeletal muscle cells from human stem cells
title_fullStr Novel integrated workflow allows production and in-depth quality assessment of multifactorial reprogrammed skeletal muscle cells from human stem cells
title_full_unstemmed Novel integrated workflow allows production and in-depth quality assessment of multifactorial reprogrammed skeletal muscle cells from human stem cells
title_short Novel integrated workflow allows production and in-depth quality assessment of multifactorial reprogrammed skeletal muscle cells from human stem cells
title_sort novel integrated workflow allows production and in-depth quality assessment of multifactorial reprogrammed skeletal muscle cells from human stem cells
topic Original Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8993739/
https://www.ncbi.nlm.nih.gov/pubmed/35396689
http://dx.doi.org/10.1007/s00018-022-04264-8
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