Department of Clinical and Molecular Medicine, Faculty of Medicine and Health Sciences, Norwegian University of Science and Technology (NTNU), PO Box 8905 MTFS, NO-7491, Trondheim, Norway.
Department of Neuromedicine and Movement Science, Faculty of Medicine and Health Sciences, Norwegian University of Science and Technology (NTNU), N-7491, Trondheim, Norway.
Biosens Bioelectron. 2019 Sep 1;140:111329. doi: 10.1016/j.bios.2019.111329. Epub 2019 May 23.
Lab-on-chip platforms, such as microfluidic chips and micro-electrode arrays (MEAs) are powerful tools that allow us to manipulate and study neurons in vitro. Microfluidic chips provide a controlled extracellular environment that structures neural networks and facilitates isolation and manipulation at a sub-cellular level. Furthermore, MEAs enable measurement of extracellular electrophysiological activity from single neurons to entire networks. Here, we demonstrate the design, fabrication and application of a 3-nodal microfluidic chip integrated with MEAs as a versatile study platform for neurobiology and pathophysiology. In this work, we evaluate the use of the microfluidic chip to structure a neural network into three separate nodes, interconnected through tunnels that isolate and guide axons into a channel, thus facilitating synaptic contacts between neurons originating from opposite nodes. Furthermore, we demonstrate the utility of the MEA for monitoring developing activity and intra-/inter nodal connectivity of the structured neural network. Finally, we demonstrate the versatility of the platform in two separate experiments. First, we demonstrate the ability to measure intra- and inter-nodal dynamic responses to a fluidically isolated chemical stimulation. Then, we demonstrate the feature of the microfluidic chip enabling the disruption of functional connectivity between nodes and examination of the immediate activity response of the neural network. The platform enables in vitro modelling of neural networks to study their functional connectomes in the context of neurodegenerative disease and CNS trauma, including spinal cord injury.
芯片实验室平台，如微流控芯片和微电极阵列（MEA），是强大的工具，使我们能够在体外操纵和研究神经元。微流控芯片提供了一个受控的细胞外环境，能够构建神经网络，并促进亚细胞水平的分离和操作。此外，MEA 能够测量从单个神经元到整个网络的细胞外电生理活性。在这里，我们展示了一种 3 节点微流控芯片与 MEA 的集成设计、制造和应用，作为神经生物学和病理生理学的多功能研究平台。在这项工作中，我们评估了微流控芯片用于将神经网络构建成三个独立节点的用途，通过隔离并引导轴突进入通道的隧道相互连接，从而促进来自相反节点的神经元之间的突触接触。此外，我们展示了 MEA 用于监测结构化神经网络的发育活动和内/节点间连接的实用性。最后，我们在两个独立的实验中展示了该平台的多功能性。首先，我们证明了该平台能够测量流体隔离化学刺激引起的节点内和节点间的动态反应。然后，我们展示了微流控芯片的功能，能够破坏节点之间的功能连接，并检查神经网络的即时活动反应。该平台能够在体外对神经网络进行建模，以研究神经退行性疾病和中枢神经系统创伤（包括脊髓损伤）背景下的功能连接组。
