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

Tomohiko Akiyama, Saeko Sato, Nana Chikazawa-Nohtomi, Atsumi Soma, Hiromi Kimura, Shunichi Wakabayashi, Shigeru Ko, Minoru Ko

Research output: Contribution to journalArticle

3 Citations (Scopus)

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.

Original languageEnglish
Article number1189
JournalScientific Reports
Volume8
Issue number1
DOIs
Publication statusPublished - 2018 Dec 1

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Pluripotent Stem Cells
Muscle Cells
Skeletal Muscle
RNA
Gene Knockdown Techniques
High-Throughput Nucleotide Sequencing
Sarcomeres
Regenerative Medicine
Muscular Dystrophies
Skeletal Muscle Fibers
Gene Expression Profiling
Muscular Diseases
Drug Discovery
Small Interfering RNA
Transcriptional Activation
Extracellular Matrix
Up-Regulation
Messenger RNA
Pharmaceutical Preparations
Therapeutics

ASJC Scopus subject areas

  • General

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Efficient differentiation of human pluripotent stem cells into skeletal muscle cells by combining RNA-based MYOD1-expression and POU5F1-silencing. / Akiyama, Tomohiko; Sato, Saeko; Chikazawa-Nohtomi, Nana; Soma, Atsumi; Kimura, Hiromi; Wakabayashi, Shunichi; Ko, Shigeru; Ko, Minoru.

In: Scientific Reports, Vol. 8, No. 1, 1189, 01.12.2018.

Research output: Contribution to journalArticle

Akiyama, Tomohiko ; Sato, Saeko ; Chikazawa-Nohtomi, Nana ; Soma, Atsumi ; Kimura, Hiromi ; Wakabayashi, Shunichi ; Ko, Shigeru ; Ko, Minoru. / Efficient differentiation of human pluripotent stem cells into skeletal muscle cells by combining RNA-based MYOD1-expression and POU5F1-silencing. In: Scientific Reports. 2018 ; Vol. 8, No. 1.
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