A Multi-Well Human iPSC Neuromuscular Disease Model on Elastomer Nanofibers

运动神经元连接中枢神经系统和肌肉，但这些突触连接的发育和动态平衡在体外很难研究，因为收缩肌纤维在标准培养系统中是不稳定的。我们建议建立一个人类IPSC神经肌肉电路模型，在该模型中，合成弹性体纳米纤维支持肌纤维的长期稳定性、排列和收缩，由96孔板中的运动神经元支配。对运动神经元的光遗传刺激将引起肌纤维收缩，提供神经肌肉传递的功能读数。高争议成像用于量化轴突、肌纤维和神经肌肉突触。该模型将通过携带影响神经肌肉连接的突变的电路进行测试。

Motor neurons connect the CNS to muscle, yet the development and homeostasis ofthese synaptic connection is difficult to study in vitro, because contractile myofibersare unstable in standard culture systems. We propose to develop a human iPSCneuromuscularcircuit model, in which synthetic elastomer nanofibers support longtermstability, alignment and contraction of myofibers, innervated by motor neurons in96-well plates. Optogenetic stimulation of motor neurons will elicit myofibercontractions, providing a functional readout of neuromuscular transmission. Highcontentimaging is used to quantify axons, myofibers, and neuromuscular synapses.The model will be tested with circuits carrying mutations that affect nerve-muscleconnectivity.