There are currently no functional neuromuscular junction (hNMJ) systems composed of human cells that could be used for drug evaluations or toxicity testing in vitro. These systems are needed to evaluate NMJs for diseases such as amyotrophic lateral sclerosis, spinal muscular atrophy or other neurodegenerative diseases or injury states. There are certainly no model systems, animal or human, that allows for isolated treatment of motoneurons or muscle capable of generating dose response curves to evaluate pharmacological activity of these highly specialized functional units. A system was developed in which human myotubes and motoneurons derived from stem cells were cultured in a serum-free medium in a BioMEMS construct. The system is composed of two chambers linked by microtunnels to enable axonal outgrowth to the muscle chamber that allows separate stimulation of each component and physiological NMJ function and MN stimulated tetanus. The muscle's contractions, induced by motoneuron activation or direct electrical stimulation, were monitored by image subtraction video recording for both frequency and amplitude. Bungarotoxin, BOTOX^® and curare dose response curves were generated to demonstrate pharmacological relevance of the phenotypic screening device. This quantifiable functional hNMJ system establishes a platform for generating patient-specific NMJ models by including patient-derived iPSCs.

目前尚无由人体细胞组成的功能性神经肌肉接头（hNMJ）系统可用于体外药物评价或毒性测试。需要这些系统来评估 NMJ 是否患有肌萎缩侧索硬化症、脊髓性肌萎缩症或其他神经退行性疾病或损伤状态等疾病。当然，没有任何动物或人类模型系统可以对运动神经元或肌肉进行单独治疗，从而生成剂量反应曲线来评估这些高度专业化的功能单元的药理活性。开发了一种系统，其中源自干细胞的人类肌管和运动神经元在 BioMEMS 结构的无血清培养基中培养。该系统由两个通过微通道连接的室组成，使轴突能够生长到肌肉室，从而可以单独刺激每个组件和生理 NMJ 功能以及 MN 刺激的破伤风。通过图像减影视频记录来监测由运动神经元激活或直接电刺激引起的肌肉收缩的频率和幅度。生成金环蛇毒素、BOTOX ^®和箭毒剂量反应曲线以证明表型筛选装置的药理学相关性。这种可量化的功能性 hNMJ 系统建立了一个平台，通过包含患者来源的 iPSC 来生成患者特异性 NMJ 模型。