title: Efficient and reproducible myogenic differentiation from human iPS cells: prospects for modeling Miyoshi Myopathy in vitro.
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.
authors.affiliation: Center for iPS Cell Research and Application (CiRA), Kyoto University, Sakyo-ku, Kyoto, Japan.
authors.forename: Akihito
authors.initials: A
authors.lastname: Tanaka
authors.forename: Knut
authors.initials: K
authors.lastname: Woltjen
authors.forename: Katsuya
authors.initials: K
authors.lastname: Miyake
authors.forename: Akitsu
authors.initials: A
authors.lastname: Hotta
authors.forename: Makoto
authors.initials: M
authors.lastname: Ikeya
authors.forename: Takuya
authors.initials: T
authors.lastname: Yamamoto
authors.forename: Tokiko
authors.initials: T
authors.lastname: Nishino
authors.forename: Emi
authors.initials: E
authors.lastname: Shoji
authors.forename: Atsuko
authors.initials: A
authors.lastname: Sehara-Fujisawa
authors.forename: Yasuko
authors.initials: Y
authors.lastname: Manabe
authors.forename: Nobuharu
authors.initials: N
authors.lastname: Fujii
authors.forename: Kazunori
authors.initials: K
authors.lastname: Hanaoka
authors.forename: Takumi
authors.initials: T
authors.lastname: Era
authors.forename: Satoshi
authors.initials: S
authors.lastname: Yamashita
authors.forename: Ken-Ichi
authors.initials: K
authors.lastname: Isobe
authors.forename: En
authors.initials: E
authors.lastname: Kimura
authors.forename: Hidetoshi
authors.initials: H
authors.lastname: Sakurai
mesh_terms: D000818:Animals; D015415:Biomarkers / Q000378:metabolism; D002454:Cell Differentiation / Q000187:drug effects* / Q000235:genetics; D002478:Cells, Cultured; D049310:Distal Myopathies / Q000235:genetics* / Q000378:metabolism / Q000473:pathology; D004318:Doxycycline / Q000494:pharmacology; D000073939:Dysferlin; D004558:Electric Stimulation; D015870:Gene Expression; D020869:Gene Expression Profiling; D005822:Genetic Vectors; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D008565:Membrane Proteins / Q000235:genetics* / Q000378:metabolism; D051379:Mice; D016513:Mice, SCID; D008954:Models, Biological; D018485:Muscle Fibers, Skeletal / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D009124:Muscle Proteins / Q000235:genetics* / Q000378:metabolism; D009133:Muscular Atrophy / Q000235:genetics* / Q000378:metabolism / Q000473:pathology; D017570:MyoD Protein / Q000235:genetics* / Q000378:metabolism; D014162:Transfection
chemical_list: D015415:Biomarkers; C114839:DYSF protein, human; D000073939:Dysferlin; D008565:Membrane Proteins; D009124:Muscle Proteins; D017570:MyoD Protein; C057363:MyoD1 myogenic differentiation protein; D004318:Doxycycline