Efficient and reproducible myogenic differentiation from human iPS cells: prospects for modeling Miyoshi Myopathy in vitro.
Efficient and reproducible myogenic differentiation from human iPS cells: prospects for modeling Miyoshi Myopathy in vitro.
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DOI：
10.1371/journal.pone.0061540
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发表时间：
2013
期刊：
影响因子：
3.7
通讯作者：
Sakurai H
中科院分区：
文献类型：
--
作者：
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
人诱导多能干细胞（hiPSC）的建立使得能够产生人类疾病的体外患者特异性细胞模型。来自患者来源的hiPSC的疾病病理学的体外重建取决于产生相关成体细胞类型的有效分化方案。然而，hiPSC的肌源性分化面临障碍，即低效率和/或差的再现性。在这里，我们报告了hiPSC快速，有效和可重复的分化为成熟的肌细胞。我们证明了在未成熟hiPSC中诱导表达肌源性分化1（MYOD 1）至少5天，驱动细胞沿着肌源性谱系沿着，效率达到70- 90%。由MYOD 1驱动的成肌分化甚至发生在未成熟的、几乎完全未分化的hiPSC中，而没有中胚层过渡。以这种方式诱导的肌细胞在分化的2周内达到成熟，如通过标记基因表达和功能特性（包括体外和体内细胞融合和响应于电刺激的抽搐）评估的。Miyoshi Myopathy（MM）是一种先天性远端肌病，由DYSFERLIN突变引起的肌膜修复缺陷引起。使用我们的诱导分化技术，我们成功地在体外重建了MM的病理状态，证明了来自MM患者的hiPSC衍生的肌管中的膜修复缺陷和通过表达全长DYSFERLIN（DYSF）的表型拯救。这些发现不仅有助于MM的病理学研究，而且可能通过使用患者来源的hiPSC用于其他人类肌肉疾病的建模。
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.
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影响因子：
6
作者：
Nagaraju, Kanneboyina;Rawat, Rashmi;Hoffman, Eric P.
通讯作者：
Hoffman, Eric P.
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通讯作者：
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通讯作者：
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作者：
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通讯作者：
Papaioannou, VE