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首页> 美国卫生研究院文献>Frontiers in Neuroengineering >Dual-Compartment Neurofluidic System for Electrophysiological Measurements in Physically Segregated and Functionally Connected Neuronal Cell Culture
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Dual-Compartment Neurofluidic System for Electrophysiological Measurements in Physically Segregated and Functionally Connected Neuronal Cell Culture

机译：用于生理隔离和功能连接的神经元细胞培养的电生理测量的双室神经流体系统。

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摘要

We developed a dual-compartment neurofluidic system with inter-connecting microchannels to connect neurons from their respective compartments, placed on a planar microelectrode arrays. The design and development of the compartmented microfluidic device for neuronal cell culture, protocol for sustaining long-term cultures, and neurite growth through microchannels in such a closed compartment device are presented. Using electrophysiological measurements of spontaneous network activity in the compartments and selective pharmacological manipulation of cells in one compartment, the biological origin of network activity and the fluidic isolation between the compartments are demonstrated. The connectivity between neuronal populations via the microchannels and the crossing-over of neurites are verified using transfection experiments and immunofluorescence staining. In addition to the neurite cross-over to the adjacent compartment, functional connectivity between cells in both the compartments is verified using cross-correlation (CC) based techniques. Bidirectional signal propagation between the compartments is demonstrated using functional connectivity maps. CC analysis and connectivity maps demonstrate that the two neuronal populations are not only functionally connected within each compartment but also with each other and a well connected functional network was formed between the compartments despite the physical barrier introduced by the microchannels.
机译：我们开发了一种双室神经流体系统，该系统具有相互连接的微通道，以将神经元从它们各自的室中连接起来，并放置在平面微电极阵列上。介绍了用于神经元细胞培养的隔室微流体装置的设计和开发，维持长期培养的方案以及在这种封闭隔室装置中通过微通道的神经突生长。使用隔室中自发网络活动的电生理测量和对一个隔室中细胞的选择性药理操作，证明了网络活动的生物学起源和隔室之间的流体隔离。使用转染实验和免疫荧光染色验证了通过微通道的神经元种群之间的连通性和神经突的交叉。除了神经突交叉到相邻的隔室之外，两个隔室中的细胞之间的功能连通性都使用基于互相关（CC）的技术进行了验证。使用功能连接图演示了隔室之间的双向信号传播。 CC分析和连通性图表明，两个神经元群体不仅在每个隔室内功能连接，而且彼此相互连接，尽管微通道引入了物理障碍，但在隔室内仍形成了连接良好的功能网络。

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