Efficient differentiation of human pluripotent stem cells into skeletal muscle cells by combining RNA-based MYOD1-expression and POU5F1-silencing

Abstract Direct generation of skeletal muscle cells from human pluripotent stem cells (hPSCs) would be beneficial for drug testing, drug discovery, and disease modelling in vitro. Here we show a rapid and robust method to induce myogenic differentiation of hPSCs by introducing mRNA encoding MYOD1 to...

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Autores principales:	Tomohiko Akiyama, Saeko Sato, Nana Chikazawa-Nohtomi, Atsumi Soma, Hiromi Kimura, Shunichi Wakabayashi, Shigeru B. H. Ko, Minoru S. H. Ko
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2018
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/5d62cce54ee541739436beee7b392604
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spelling	oai:doaj.org-article:5d62cce54ee541739436beee7b3926042021-12-02T15:09:03ZEfficient differentiation of human pluripotent stem cells into skeletal muscle cells by combining RNA-based MYOD1-expression and POU5F1-silencing10.1038/s41598-017-19114-y2045-2322https://doaj.org/article/5d62cce54ee541739436beee7b3926042018-01-01T00:00:00Zhttps://doi.org/10.1038/s41598-017-19114-yhttps://doaj.org/toc/2045-2322Abstract Direct generation of skeletal muscle cells from human pluripotent stem cells (hPSCs) would be beneficial for drug testing, drug discovery, and disease modelling in vitro. Here we show a rapid and robust method to induce myogenic differentiation of hPSCs by introducing mRNA encoding MYOD1 together with siRNA-mediated knockdown of POU5F1 (also known as OCT4 or OCT3/4). This integration-free approach generates functional skeletal myotubes with sarcomere-like structure and a fusion capacity in several days. The POU5F1 silencing facilitates MYOD1 recruitment to the target promoters, which results in the significant activation of myogenic genes in hPSCs. Furthermore, deep sequencing transcriptome analyses demonstrated that POU5F1-knockdown upregulates the genes associated with IGF- and FGF-signaling and extracellular matrix that may also support myogenic differentiation. This rapid and direct differentiation method may have potential applications in regenerative medicine and disease therapeutics for muscle disorders such as muscular dystrophy.Tomohiko AkiyamaSaeko SatoNana Chikazawa-NohtomiAtsumi SomaHiromi KimuraShunichi WakabayashiShigeru B. H. KoMinoru S. H. KoNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 8, Iss 1, Pp 1-13 (2018)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Tomohiko Akiyama Saeko Sato Nana Chikazawa-Nohtomi Atsumi Soma Hiromi Kimura Shunichi Wakabayashi Shigeru B. H. Ko Minoru S. H. Ko Efficient differentiation of human pluripotent stem cells into skeletal muscle cells by combining RNA-based MYOD1-expression and POU5F1-silencing
description	Abstract Direct generation of skeletal muscle cells from human pluripotent stem cells (hPSCs) would be beneficial for drug testing, drug discovery, and disease modelling in vitro. Here we show a rapid and robust method to induce myogenic differentiation of hPSCs by introducing mRNA encoding MYOD1 together with siRNA-mediated knockdown of POU5F1 (also known as OCT4 or OCT3/4). This integration-free approach generates functional skeletal myotubes with sarcomere-like structure and a fusion capacity in several days. The POU5F1 silencing facilitates MYOD1 recruitment to the target promoters, which results in the significant activation of myogenic genes in hPSCs. Furthermore, deep sequencing transcriptome analyses demonstrated that POU5F1-knockdown upregulates the genes associated with IGF- and FGF-signaling and extracellular matrix that may also support myogenic differentiation. This rapid and direct differentiation method may have potential applications in regenerative medicine and disease therapeutics for muscle disorders such as muscular dystrophy.
format	article
author	Tomohiko Akiyama Saeko Sato Nana Chikazawa-Nohtomi Atsumi Soma Hiromi Kimura Shunichi Wakabayashi Shigeru B. H. Ko Minoru S. H. Ko
author_facet	Tomohiko Akiyama Saeko Sato Nana Chikazawa-Nohtomi Atsumi Soma Hiromi Kimura Shunichi Wakabayashi Shigeru B. H. Ko Minoru S. H. Ko
author_sort	Tomohiko Akiyama
title	Efficient differentiation of human pluripotent stem cells into skeletal muscle cells by combining RNA-based MYOD1-expression and POU5F1-silencing
title_short	Efficient differentiation of human pluripotent stem cells into skeletal muscle cells by combining RNA-based MYOD1-expression and POU5F1-silencing
title_full	Efficient differentiation of human pluripotent stem cells into skeletal muscle cells by combining RNA-based MYOD1-expression and POU5F1-silencing
title_fullStr	Efficient differentiation of human pluripotent stem cells into skeletal muscle cells by combining RNA-based MYOD1-expression and POU5F1-silencing
title_full_unstemmed	Efficient differentiation of human pluripotent stem cells into skeletal muscle cells by combining RNA-based MYOD1-expression and POU5F1-silencing
title_sort	efficient differentiation of human pluripotent stem cells into skeletal muscle cells by combining rna-based myod1-expression and pou5f1-silencing
publisher	Nature Portfolio
publishDate	2018
url	https://doaj.org/article/5d62cce54ee541739436beee7b392604
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Efficient differentiation of human pluripotent stem cells into skeletal muscle cells by combining RNA-based MYOD1-expression and POU5F1-silencing

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