RRC ID 50084
著者 Akiyama T, Sato S, Chikazawa-Nohtomi N, Soma A, Kimura H, Wakabayashi S, Ko SBH, Ko MSH.
タイトル Efficient differentiation of human pluripotent stem cells into skeletal muscle cells by combining RNA-based MYOD1-expression and POU5F1-silencing.
ジャーナル Sci Rep
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.
巻・号 8(1)
ページ 1189
公開日 2018-1-19
DOI 10.1038/s41598-017-19114-y
PII 10.1038/s41598-017-19114-y
PMID 29352121
PMC PMC5775307
MeSH Cell Differentiation / genetics* Cell Line Embryonic Stem Cells / cytology Embryonic Stem Cells / metabolism Gene Expression Regulation Gene Knockdown Techniques Gene Silencing Humans Models, Biological Muscle Development / genetics Muscle Fibers, Skeletal / cytology* Muscle Fibers, Skeletal / metabolism* MyoD Protein / genetics* Octamer Transcription Factor-3 / genetics* Pluripotent Stem Cells / cytology* Pluripotent Stem Cells / metabolism*
IF 3.998
引用数 3
リソース情報
ヒト・動物細胞 201B7(HPS0063) 409B2(HPS0076)