3-D multi-electrode arrays detect early spontaneous electrophysiological activity in 3-D neuronal-astrocytic co-cultures.
3-D multi-electrode arrays detect early spontaneous electrophysiological activity in 3-D neuronal-astrocytic co-cultures.
复制标题
3-D 多电极阵列检测 3-D 神经元-星形细胞共培养物中的早期自发电生理活动。
DOI：
10.1007/s13534-020-00166-5
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发表时间：
2020
影响因子：
4.6
通讯作者：
LaPlaca,MichelleC
中科院分区：
文献类型：
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作者：
Vernekar,VaradrajN;LaPlaca,MichelleC
三维（3-D）神经培养代表了研究疾病和药物筛选的一个有前途的平台。需要用于测量这些培养物中的电生理功能的工具和方法。因此，这项工作的目的主要是开发一种方法，使工程3-D解离神经培养物与市售的3-D多电极阵列（MEA）可靠地在3周内，使其电生理活动的记录。我们进一步比较了这些培养物的功能输出与其结构和突触网络随时间的发展。我们可靠地将原代啮齿动物神经元-星形胶质细胞（2：1）3-D共培养物（2500个细胞/mm 3平板细胞密度）在Matrigel™（7.5 mg/mL）中与掺加的3-D MEA连接，同时保持高活力，所述Matrigel ™（7.5 mg/mL）厚达750 μm（30-40个细胞层）。使用这些MEA，我们成功地记录了神经网络水平的电生理活性的自发发展，并在体外3周内使用免疫细胞化学在这些共培养物中测量了推定的突触和神经元成熟的发展。平面（2-D）多边环境协定与这些文化的接口作为记录控制。在这个接口的3-D文化MEA系统内的神经元表现出相当大的神经突生长，网络，神经元成熟，突触，和文化范围内的自发放电的同步尖峰和动作电位的爆发。在培养的第一周期间，网络范围的尖峰和同步爆发迅速增加（在第2天首次检测到），在第二周期间达到平台，并且在第三周中略微减少，同时在整个3周培养期内保持高活力。早期电生理活动发生在神经元突起成熟之前，第二周突触密度显著增加。我们成功地将3-D神经共培养物与3-D MEA连接，并记录了这些培养物在3周内的电生理活动。在这些文化中的电生理活动的快速增加，随后由一段时间的神经元成熟和高水平的突触形成的初始阶段表明一个发展的稳态过程。这种方法可以使未来的应用在神经网络行为的基础研究和涉及药物测试和神经接口的转化研究。
Three-dimensional (3-D) neural cultures represent a promising platform for studying disease and drug screening. Tools and methodologies for measuring the electrophysiological function in these cultures are needed. Therefore, the purpose of this work was primarily to develop a methodology to interface engineered 3-D dissociated neural cultures with commercially available 3-D multi-electrode arrays (MEAs) reliably over 3 weeks to enable the recording of their electrophysiological activity. We further compared the functional output of these cultures to their structural and synaptic network development over time. We reliably interfaced a primary rodent neuron-astrocyte (2:1) 3-D co-culture (2500 cells/mm3plating cell density) in Matrigel™ (7.5 mg/mL) that was up to 750 µm thick (30–40 cell-layers) with spiked 3-D MEAs while maintaining high viability. Using these MEAs we successfully recorded the spontaneous development of neural network-level electrophysiological activity and measured the development of putative synapses and neuronal maturation in these co-cultures using immunocytochemistry over 3 weeks in vitro. Planar (2-D) MEAs interfaced with these cultures served as recording controls. Neurons within this interfaced 3-D culture-MEA system exhibited considerable neurite outgrowth, networking, neuronal maturation, synaptogenesis, and culture-wide spontaneous firing of synchronized spikes and bursts of action potentials. Network-wide spikes and synchronized bursts increased rapidly (first detected at 2 days) during the first week in culture, plateaued during the second week, and reduced slightly in the third week, while maintaining high viability throughout the 3-week culturing period. Early electrophysiology activity occurred prior to neuronal process maturation and significant synaptic density increases in the second week. We successfully interfaced 3-D neural co-cultures with 3-D MEAs and recorded the electrophysiological activity of these cultures over 3 weeks. The initial period of rapid increase in electrophysiological activity, followed by a period of neuronal maturation and high-level of synapse formation in these cultures suggests a developmental homeostatic process. This methodology can enable future applications both in fundamental investigations of neural network behavior and in translational studies involving drug testing and neural interfacing.