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  • A muscle fatigue-like contractile decline was recapitulated using skeletal myotubes from Duchenne muscular dystrophy patient-derived iPSCs.

A muscle fatigue-like contractile decline was recapitulated using skeletal myotubes from Duchenne muscular dystrophy patient-derived iPSCs.

Cell reports. Medicine (2021-07-02)
Tomoya Uchimura, Toshifumi Asano, Takao Nakata, Akitsu Hotta, Hidetoshi Sakurai
要旨

Duchenne muscular dystrophy (DMD) is a muscle degenerating disease caused by dystrophin deficiency, for which therapeutic options are limited. To facilitate drug development, it is desirable to develop in vitro disease models that enable the evaluation of DMD declines in contractile performance. Here, we show MYOD1-induced differentiation of hiPSCs into functional skeletal myotubes in vitro with collagen gel and electrical field stimulation (EFS). Long-term EFS training (0.5 Hz, 20 V, 2 ms, continuous for 2 weeks) mimicking muscle overuse recapitulates declines in contractile performance in dystrophic myotubes. A screening of clinically relevant drugs using this model detects three compounds that ameliorate this decline. Furthermore, we validate the feasibility of adapting the model to a 96-well culture system using optogenetic technology for large-scale screening. Our results support a disease model using patient-derived iPSCs that allows for the recapitulation of the contractile pathogenesis of DMD and a screening strategy for drug development.

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Sigma-Aldrich
ピューロマイシン 二塩酸塩 Streptomyces alboniger由来, powder, BioReagent, suitable for cell culture
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レチノイン酸, ≥98% (HPLC), powder
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3,3′,5-トリヨード-L-チロニン ナトリウム塩, powder, BioReagent, suitable for cell culture
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ウマ血清, Donor Herd, USA origin, Heat inactivated, sterile-filtered, suitable for cell culture
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モノクロナール抗ミオシン (骨格筋, 速筋型) マウス宿主抗体, clone MY-32, ascites fluid
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Anti-Myosin-2 (MYH2) Antibody, clone SC-71, clone SC-71, from mouse