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) |