RRC ID 44416
著者 Tanaka A, Woltjen K, Miyake K, Hotta A, Ikeya M, Yamamoto T, Nishino T, Shoji E, Sehara-Fujisawa A, Manabe Y, Fujii N, Hanaoka K, Era T, Yamashita S, Isobe K, Kimura E, Sakurai H.
タイトル Efficient and reproducible myogenic differentiation from human iPS cells: prospects for modeling Miyoshi Myopathy in vitro.
ジャーナル PLoS One
Abstract The establishment of human induced pluripotent stem cells (hiPSCs) has enabled the production of in vitro, patient-specific cell models of human disease. In vitro recreation of disease pathology from patient-derived hiPSCs depends on efficient differentiation protocols producing relevant adult cell types. However, myogenic differentiation of hiPSCs has faced obstacles, namely, low efficiency and/or poor reproducibility. Here, we report the rapid, efficient, and reproducible differentiation of hiPSCs into mature myocytes. We demonstrated that inducible expression of myogenic differentiation1 (MYOD1) in immature hiPSCs for at least 5 days drives cells along the myogenic lineage, with efficiencies reaching 70-90%. Myogenic differentiation driven by MYOD1 occurred even in immature, almost completely undifferentiated hiPSCs, without mesodermal transition. Myocytes induced in this manner reach maturity within 2 weeks of differentiation as assessed by marker gene expression and functional properties, including in vitro and in vivo cell fusion and twitching in response to electrical stimulation. Miyoshi Myopathy (MM) is a congenital distal myopathy caused by defective muscle membrane repair due to mutations in DYSFERLIN. Using our induced differentiation technique, we successfully recreated the pathological condition of MM in vitro, demonstrating defective membrane repair in hiPSC-derived myotubes from an MM patient and phenotypic rescue by expression of full-length DYSFERLIN (DYSF). These findings not only facilitate the pathological investigation of MM, but could potentially be applied in modeling of other human muscular diseases by using patient-derived hiPSCs.
巻・号 8(4)
ページ e61540
公開日 2013-1-1
DOI 10.1371/journal.pone.0061540
PII PONE-D-12-35707
PMID 23626698
PMC PMC3633995
MeSH Animals Biomarkers / metabolism Cell Differentiation / drug effects* Cell Differentiation / genetics Cells, Cultured Distal Myopathies / genetics* Distal Myopathies / metabolism Distal Myopathies / pathology Doxycycline / pharmacology Dysferlin Electric Stimulation Gene Expression Gene Expression Profiling Genetic Vectors Humans Induced Pluripotent Stem Cells / cytology Induced Pluripotent Stem Cells / drug effects Induced Pluripotent Stem Cells / metabolism* Membrane Proteins / genetics* Membrane Proteins / metabolism Mice Mice, SCID Models, Biological Muscle Fibers, Skeletal / cytology Muscle Fibers, Skeletal / drug effects Muscle Fibers, Skeletal / metabolism* Muscle Proteins / genetics* Muscle Proteins / metabolism Muscular Atrophy / genetics* Muscular Atrophy / metabolism Muscular Atrophy / pathology MyoD Protein / genetics* MyoD Protein / metabolism Transfection
IF 2.74
引用数 99
WOS 分野 MULTIDISCIPLINARY SCIENCES
リソース情報
ヒト・動物細胞 Hu5/E18(RCB2366)