Abstract

3D bioengineered skeletal muscle macrotissues are increasingly important for studies of cell biology and development of therapeutics. Tissues derived from immortalized cells obtained from patient samples, or from pluripotent stem cells, can be co-cultured with motor-neurons to create models of human neuromuscular junctions in culture. In this study, we present foundational work on 3D cultured muscle ultrastructure, with and without motor neurons, which is enabled by the development of a new co-culture platform. Our results show that tissues from Duchenne muscular dystrophy patients are poorly organized compared to tissues grown from healthy donor and that the presence of motor neurons invariably improves sarcomere organization. Electron micrographs show that in the presence of motor neurons, filament directionality, banding patterns, z-disc continuity, and the appearance of presumptive SSR and T-tubule profiles all improve in healthy, DMD-, and iPSC-derived muscle tissue. Further work to identify the underlying defects of DMD tissue disorganization and the mechanisms by which motor neurons support muscle are likely to yield potential new therapeutic approaches for treating patients suffering from Duchenne muscular dystrophy.

中文翻译：

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电子显微镜分析 iPSC 衍生的运动神经元对生物工程人体骨骼肌组织的影响

摘要

3D 生物工程骨骼肌大组织对于细胞生物学研究和治疗方法的开发越来越重要。来自患者样本或多能干细胞的永生化细胞的组织可以与运动神经元共培养，以创建人类神经肌肉接头的培养模型。在这项研究中，我们展示了 3D 培养肌肉超微结构（有或没有运动神经元）的基础工作，这是通过开发新的共培养平台实现的。我们的结果表明，与健康供体生长的组织相比，杜氏肌营养不良症患者的组织组织较差，并且运动神经元的存在总是会改善肌节组织。电子显微照片显示，在运动神经元存在的情况下，健康的 DMD 和 iPSC 衍生的肌肉组织中的肌丝方向性、条带模式、z 盘连续性以及假定的 SSR 和 T 管轮廓的出现均得到改善。进一步研究确定 DMD 组织紊乱的潜在缺陷以及运动神经元支持肌肉的机制可能会产生治疗杜氏肌营养不良症患者的潜在新治疗方法。