Multilineage Differentiation for Formation of Innervated Skeletal Muscle Fibers from Healthy and Diseased Human Pluripotent Stem Cells
Induced pluripotent stem cells (iPSCs) obtained by reprogramming primary somatic cells have revolutionized the fields of cell biology and disease modeling. However, the number protocols for generating mature muscle fibers with sarcolemmal organization using iPSCs remain limited, and partly mimic the...
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doaj-8d061bb2ffea436e998913b85cd1c2af2020-11-25T03:02:24ZengMDPI AGCells2073-44092020-06-0191531153110.3390/cells9061531Multilineage Differentiation for Formation of Innervated Skeletal Muscle Fibers from Healthy and Diseased Human Pluripotent Stem CellsKilian Mazaleyrat0Cherif Badja1Natacha Broucqsault2Raphaël Chevalier3Camille Laberthonnière4Camille Dion5Lyla Baldasseroni6Claire El-Yazidi7Morgane Thomas8Richard Bachelier9Alexandre Altié10Karine Nguyen11Nicolas Lévy12Jérôme D. Robin13Frédérique Magdinier14Aix-Marseille University, INSERM, MMG, Marseille Medical Genetics, 13385 Marseille, FranceAix-Marseille University, INSERM, MMG, Marseille Medical Genetics, 13385 Marseille, FranceAix-Marseille University, INSERM, MMG, Marseille Medical Genetics, 13385 Marseille, FranceAix-Marseille University, INSERM, MMG, Marseille Medical Genetics, 13385 Marseille, FranceAix-Marseille University, INSERM, MMG, Marseille Medical Genetics, 13385 Marseille, FranceAix-Marseille University, INSERM, MMG, Marseille Medical Genetics, 13385 Marseille, FranceAix-Marseille University, INSERM, MMG, Marseille Medical Genetics, 13385 Marseille, FranceAix-Marseille University, INSERM, MMG, Marseille Medical Genetics, 13385 Marseille, FranceAix-Marseille University, INSERM, MMG, Marseille Medical Genetics, 13385 Marseille, FranceAix-Marseille University, INSERM, INRA, C2VN, 13385 Marseille, FranceAix-Marseille University, INSERM, INRA, C2VN, 13385 Marseille, FranceAix-Marseille University, INSERM, MMG, Marseille Medical Genetics, 13385 Marseille, FranceAix-Marseille University, INSERM, MMG, Marseille Medical Genetics, 13385 Marseille, FranceAix-Marseille University, INSERM, MMG, Marseille Medical Genetics, 13385 Marseille, FranceAix-Marseille University, INSERM, MMG, Marseille Medical Genetics, 13385 Marseille, FranceInduced pluripotent stem cells (iPSCs) obtained by reprogramming primary somatic cells have revolutionized the fields of cell biology and disease modeling. However, the number protocols for generating mature muscle fibers with sarcolemmal organization using iPSCs remain limited, and partly mimic the complexity of mature skeletal muscle. <b>Methods:</b> We used a novel combination of small molecules added in a precise sequence for the simultaneous codifferentiation of human iPSCs into skeletal muscle cells and motor neurons. <b>Results:</b> We show that the presence of both cell types reduces the production time for millimeter-long multinucleated muscle fibers with sarcolemmal organization. Muscle fiber contractions are visible in 19–21 days, and can be maintained over long period thanks to the production of innervated multinucleated mature skeletal muscle fibers with autonomous cell regeneration of PAX7-positive cells and extracellular matrix synthesis. The sequential addition of specific molecules recapitulates key steps of human peripheral neurogenesis and myogenesis. Furthermore, this organoid-like culture can be used for functional evaluation and drug screening. <b>Conclusion:</b> Our protocol, which is applicable to hiPSCs from healthy individuals, was validated in Duchenne Muscular Dystrophy, Myotonic Dystrophy, Facio-Scapulo-Humeral Dystrophy and type 2A Limb-Girdle Muscular Dystrophy, opening new paths for the exploration of muscle differentiation, disease modeling and drug discovery.https://www.mdpi.com/2073-4409/9/6/1531human induced pluripotent cellsdifferentiationmyoblastsmyotubesmotor neuronssatellite cells |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Kilian Mazaleyrat Cherif Badja Natacha Broucqsault Raphaël Chevalier Camille Laberthonnière Camille Dion Lyla Baldasseroni Claire El-Yazidi Morgane Thomas Richard Bachelier Alexandre Altié Karine Nguyen Nicolas Lévy Jérôme D. Robin Frédérique Magdinier |
spellingShingle |
Kilian Mazaleyrat Cherif Badja Natacha Broucqsault Raphaël Chevalier Camille Laberthonnière Camille Dion Lyla Baldasseroni Claire El-Yazidi Morgane Thomas Richard Bachelier Alexandre Altié Karine Nguyen Nicolas Lévy Jérôme D. Robin Frédérique Magdinier Multilineage Differentiation for Formation of Innervated Skeletal Muscle Fibers from Healthy and Diseased Human Pluripotent Stem Cells Cells human induced pluripotent cells differentiation myoblasts myotubes motor neurons satellite cells |
author_facet |
Kilian Mazaleyrat Cherif Badja Natacha Broucqsault Raphaël Chevalier Camille Laberthonnière Camille Dion Lyla Baldasseroni Claire El-Yazidi Morgane Thomas Richard Bachelier Alexandre Altié Karine Nguyen Nicolas Lévy Jérôme D. Robin Frédérique Magdinier |
author_sort |
Kilian Mazaleyrat |
title |
Multilineage Differentiation for Formation of Innervated Skeletal Muscle Fibers from Healthy and Diseased Human Pluripotent Stem Cells |
title_short |
Multilineage Differentiation for Formation of Innervated Skeletal Muscle Fibers from Healthy and Diseased Human Pluripotent Stem Cells |
title_full |
Multilineage Differentiation for Formation of Innervated Skeletal Muscle Fibers from Healthy and Diseased Human Pluripotent Stem Cells |
title_fullStr |
Multilineage Differentiation for Formation of Innervated Skeletal Muscle Fibers from Healthy and Diseased Human Pluripotent Stem Cells |
title_full_unstemmed |
Multilineage Differentiation for Formation of Innervated Skeletal Muscle Fibers from Healthy and Diseased Human Pluripotent Stem Cells |
title_sort |
multilineage differentiation for formation of innervated skeletal muscle fibers from healthy and diseased human pluripotent stem cells |
publisher |
MDPI AG |
series |
Cells |
issn |
2073-4409 |
publishDate |
2020-06-01 |
description |
Induced pluripotent stem cells (iPSCs) obtained by reprogramming primary somatic cells have revolutionized the fields of cell biology and disease modeling. However, the number protocols for generating mature muscle fibers with sarcolemmal organization using iPSCs remain limited, and partly mimic the complexity of mature skeletal muscle. <b>Methods:</b> We used a novel combination of small molecules added in a precise sequence for the simultaneous codifferentiation of human iPSCs into skeletal muscle cells and motor neurons. <b>Results:</b> We show that the presence of both cell types reduces the production time for millimeter-long multinucleated muscle fibers with sarcolemmal organization. Muscle fiber contractions are visible in 19–21 days, and can be maintained over long period thanks to the production of innervated multinucleated mature skeletal muscle fibers with autonomous cell regeneration of PAX7-positive cells and extracellular matrix synthesis. The sequential addition of specific molecules recapitulates key steps of human peripheral neurogenesis and myogenesis. Furthermore, this organoid-like culture can be used for functional evaluation and drug screening. <b>Conclusion:</b> Our protocol, which is applicable to hiPSCs from healthy individuals, was validated in Duchenne Muscular Dystrophy, Myotonic Dystrophy, Facio-Scapulo-Humeral Dystrophy and type 2A Limb-Girdle Muscular Dystrophy, opening new paths for the exploration of muscle differentiation, disease modeling and drug discovery. |
topic |
human induced pluripotent cells differentiation myoblasts myotubes motor neurons satellite cells |
url |
https://www.mdpi.com/2073-4409/9/6/1531 |
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