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Abstract：
Engineered 3D neural tissues made of neurons and glial cells derived from human induced pluripotent stem cells (hiPSC) are among the most promising tools in drug discovery and neurotoxicology. They represent a cheaper, faster, and more ethical alternative to in vivo animal testing that will likely close the gap between in vitro animal models and human clinical trials. Micro-Electrode Array (MEA) technology is known to provide an assessment of compound effects on neural 2D cell cultures and acute tissue preparations by real-time, non-invasive, and long-lasting electrophysiological monitoring of spontaneous and evoked neuronal activity. Nevertheless, the use of engineered 3D neural tissues in combination with MEA biochips still involves series of constraints, such as drastically limited diffusion of oxygen and nutrients within tissues mainly due to the lack of vascularization. Therefore, 3D neural tissues are extremely sensitive to experimental conditions and require an adequately designed interface that provides optimal tissue survival conditions. A well-suited technique to overcome this issue is the combination of the Air-Liquid Interface (ALI) tissue culture method with the MEA technology. We have developed a full 3D neural tissue culture process and a data acquisition system composed of high-end electronics and novel MEA biochips based on porous, flexible, thin-film membranes integrating recording electrodes, named as \"Strip-MEA,\" to allow the maintenance of an ALI around the 3D neural tissues. The main motivation of the porous MEA biochips development was the possibility to monitor and to study the electrical activity of 3D neural tissues under different recording configurations, (i) the Strip-MEA can be placed below a tissue, (ii) or by taking advantage of the ALI, be directly placed on top of the tissue, or finally, (iii) it can be embedded into a larger neural tissue generated by the fusion of two (or more) tissues placed on both sides of the Strip-MEA allowing the recording from its inner part. This paper presents the recording and analyses of spontaneous activity from the three positioning configurations of the Strip-MEAs. Obtained results are discussed with the perspective of developing in vitro models of brain diseases and/or impairment of neural network functioning.
摘要：
由源自人诱导多能干细胞（hiPSC）的神经元和神经胶质细胞制成的工程化3D神经组织是药物发现和神经毒理学中最有前途的工具之一。他们代表一个更便宜的，更快,和更多的伦理替代体内动物试验，这将可能缩小体外动物模型和人体临床试验之间的差距。众所周知，微电极阵列（MEA）技术可以实时评估复合对神经2D细胞培养物和急性组织制剂的影响。非侵入性，以及对自发和诱发的神经元活动的长期电生理监测。然而，工程3D神经组织与MEA生物芯片的结合使用仍然涉及一系列限制，例如，主要由于缺乏血管形成，氧气和营养物质在组织内的扩散受到极大限制。因此,3D神经组织对实验条件极其敏感,并且需要提供最佳组织存活条件的适当设计的接口。克服该问题的一种非常适合的技术是空气-液体界面(ALI)组织培养方法与MEA技术的组合。我们开发了一个完整的3D神经组织培养过程和一个数据采集系统，该系统由高端电子设备和基于多孔，灵活,集成记录电极的薄膜膜，命名为“Strip-MEA，“允许在3D神经组织周围维持ALI。多孔MEA生物芯片开发的主要动机是在不同记录配置下监测和研究3D神经组织的电活动的可能性。(i)条带-MEA可以放置在纸巾下面，（ii）或利用ALI，直接放在组织的顶部，或者最后，(iii)它可以被嵌入到较大的神经组织中,该较大的神经组织是由放置在条带-MEA两侧的两个(或更多个)组织的融合产生的,从而允许从其内部记录。本文介绍了从Strip-MEAs的三种定位配置对自发活动的记录和分析。从开发脑疾病和/或神经网络功能受损的体外模型的角度讨论了获得的结果。
