In Vitro Evaluation of Exon Skipping in Disease-Specific iPSC-Derived Myocytes
In Vitro Evaluation of Exon Skipping in Disease-Specific iPSC-Derived Myocytes
复制标题
疾病特异性 iPSC 衍生肌细胞中外显子跳跃的体外评估
DOI：
10.1007/978-1-4939-8651-4_11
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发表时间：
2018
期刊：
影响因子：
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通讯作者：
Hidetoshi Sakurai
中科院分区：
文献类型：
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作者：
Mingming Zhao;Emi Shoji;Hidetoshi Sakurai
患者来源的疾病特异性诱导多能干细胞(IPSCs)已经打开了体外重建病理条件的大门，通过分化成与每个靶组织相对应的疾病细胞。为了研究肌肉疾病，我们建立了一种通过诱导MYOD1表达来诱导人iPSCs进入肌细胞的成肌分化方案。这种高度可重复性的分化方案产生了同质的骨骼肌细胞群，效率高达70%-90%。如此高的效率使我们能够评估疾病特异性心肌细胞中外显子跳跃的有效性。这些疾病特异性IPSC来源的心肌细胞不仅可用于验证特定反义寡核苷酸的治疗效果，还可用于结合多孔分化系统筛选跳过外显子的化学物质。
Patient-derived disease-specific induced pluripotent stem cells (iPSCs) have opened the door to recreating pathological conditions in vitro using differentiation into diseased cells corresponding to each target tissue. To investigate muscular disease, we have established a myogenic differentiation protocol mediated by inducibleMYOD1expression that drives human iPSCs into myocytes. This highly reproducible differentiation protocol yields a homogenous skeletal muscle cell population, reaching efficiencies as high as 70–90%. Such high efficiency enables us to evaluate the efficacy of exon skipping in disease-specific myocytes. These disease-specific iPSC-derived myocytes can be applied not only for the validation of therapeutic efficacy of specific antisense oligonucleotide but also for the screening of exon skipping chemicals combined with the multiwell differentiation system.