Hybrid Systems Lab, NanoScience Technology Center, University of Central Florida, 12424 Research Parkway, Suite 400, Orlando, FL 32826, USA.
Hybrid Systems Lab, NanoScience Technology Center, University of Central Florida, 12424 Research Parkway, Suite 400, Orlando, FL 32826, USA; Department of Biomedical Engineering, 305 Weill Hall, Cornell University, Ithaca, NY 14853, USA.
Biomaterials. 2018 Jun;166:64-78. doi: 10.1016/j.biomaterials.2018.02.047. Epub 2018 Feb 27.
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，可用于体外药物评估或毒性测试。这些系统对于评估肌萎缩侧索硬化症、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的 NMJs 是必需的。当然，没有任何模型系统，无论是动物还是人类，都可以单独治疗运动神经元或肌肉，以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学活性。开发了一种系统，其中源自干细胞的人肌管和运动神经元在无血清培养基中在 BioMEMS 构建体中培养。该系统由两个腔室组成，通过微隧道连接，以允许轴突生长到肌肉腔室，从而能够单独刺激每个组件和生理 NMJ 功能以及 MN 刺激的强直。通过图像相减视频记录监测肌肉的收缩，以监测频率和幅度。生成了 Bungarotoxin、BOTOX 和 curare 的剂量反应曲线，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 系统通过包含患者来源的 iPSCs，为生成患者特异性 NMJ 模型建立了一个平台。
