This protocol describes the design, fabrication and use of a 3D physiological and pathophysiological motor unit model consisting of motor neurons coupled to skeletal muscles interacting via the neuromuscular junction (NMJ) within a microfluidic device. This model facilitates imaging and quantitative functional assessment. The ‘NMJ chip’ enables real-time, live imaging of axonal outgrowth, NMJ formation and muscle maturation, as well as synchronization of motor neuron activity and muscle contraction under optogenetic control for the study of normal physiological events. The proposed protocol takes ~2–3 months to be implemented. Pathological behaviors associated with various neuromuscular diseases, such as regression of motor neuron axons, motor neuron death, and muscle degradation and atrophy can also be recapitulated in this system. Disease models can be created by the use of patient-derived induced pluripotent stem cells to generate both the motor neurons and skeletal muscle cells used. This is demonstrated by the use of cells from a patient with sporadic amyotrophic lateral sclerosis but can be applied more generally to models of neuromuscular disease, such as spinal muscular atrophy, NMJ disorder and muscular dystrophy. Models such as this hold considerable potential for applications in precision medicine, drug screening and disease risk assessment.

该协议描述了 3D 生理和病理生理运动单元模型的设计、制造和使用，该模型由运动神经元耦合到通过微流体装置内的神经肌肉接头 (NMJ) 相互作用的骨骼肌。该模型有助于成像和定量功能评估。“NMJ 芯片”可实现轴突生长、NMJ 形成和肌肉成熟的实时实时成像，以及在光遗传学控制下同步运动神经元活动和肌肉收缩，以研究正常生理事件。拟议的协议需要约 2-3 个月才能实施。与各种神经肌肉疾病相关的病理行为，如运动神经元轴突退化、运动神经元死亡、肌肉退化和萎缩等，也可以在该系统中得到概括。疾病模型可以通过使用源自患者的诱导多能干细胞来生成所使用的运动神经元和骨骼肌细胞来创建。使用来自散发性肌萎缩侧索硬化症患者的细胞证明了这一点，但可以更普遍地应用于神经肌肉疾病模型，例如脊髓性肌萎缩症、NMJ 疾病和肌营养不良症。诸如此类的模型在精准医学、药物筛选和疾病风险评估中具有相当大的应用潜力。如脊髓性肌萎缩症、NMJ 障碍和肌营养不良症。诸如此类的模型在精准医学、药物筛选和疾病风险评估中具有相当大的应用潜力。如脊髓性肌萎缩症、NMJ 障碍和肌营养不良症。诸如此类的模型在精准医学、药物筛选和疾病风险评估中具有相当大的应用潜力。