A high-throughput spheroid fusion platform for the templated-assembly of 3D neuromuscular junctions

神经元通过从大脑和脊髓向肌肉发送电脉冲来帮助控制运动。神经元和肌肉之间的连接发生在人体的特定部位，称为神经肌肉连接点（NMJ）。了解神经元和肌肉细胞之间的相互作用对于理解神经肌肉的发育和功能障碍至关重要。目前，还没有可用的人体三维模型来研究这种连接，因此通常使用动物模型来代替。然而，动物的NMJ与人类的NMJ有很大的不同。因此，开发准确的NMJ人体模型至关重要。在过去的几年里，我们的团队开发了新的技术，允许实验室培养组织的形成，可以用来概括人类生理。在本项目中，我们将创建一种新的体外神经肌肉连接模型，以减轻对动物模型的需求。我们将把神经元和肌肉细胞都培养成三维培养物，并利用微陷阱网络迫使它们组装起来。我们将首次生成真正的3D类器官，每个实验都有数百个这样的连接，这个数字至少比以前的尝试高出一个数量级。这些连接将通过测量神经元受到化学刺激时的肌肉收缩来验证。这将确保我们创建的模型与人类生理学具有最高的相关性，并且可以用作未来生物医学研究的平台。

Neurons help control movement by sending conveying electrical impulses from the brain and spinal cord to the muscles. The connection between the neurons and muscles happens in the human body at specific places called neuromuscular junctions (NMJ). Understanding the interactions between neurons and muscle cells is crucial to understand neuromuscular development and dysfunction. At the moment, there are no available human three-dimensional models that can be used to study this junction, hence animal models are routinely used instead. However, animal NMJ differs significantly from the human ones. Hence, it is critical to develop accurate human models of the NMJ. In the past few years, our team has developed novel technologies that permit the formation of lab-grown tissue that can be used to recapitulate human physiology.In this project, we will create a novel in-vitro model neuromuscular junction to alleviate the need for animal models. We will grow both neurons and muscle cells into three dimensional cultures and force them to assemble using networks of microtraps. We will generate for the first time truly 3D organoids with hundreds of such junctions per experiment, a number at least one order of magnitude higher than previous attempts. The junctions will be validated by measuring muscle contraction upon chemical stimulation of the neurons. This will ensure that the models we create have the highest relevance to human physiology and can be used as a platform for future biomedical research.