Physical and Life Sciences Directorate, Lawrence Livermore National Laboratory, Livermore, CA, USA.
Engineering Directorate, Lawrence Livermore National Laboratory, Livermore, CA, USA.
Sci Rep. 2020 Jul 3;10(1):11007. doi: 10.1038/s41598-020-67691-2.
Brain-on-a-chip systems are designed to simulate brain activity using traditional in vitro cell culture on an engineered platform. It is a noninvasive tool to screen new drugs, evaluate toxicants, and elucidate disease mechanisms. However, successful recapitulation of brain function on these systems is dependent on the complexity of the cell culture. In this study, we increased cellular complexity of traditional (simple) neuronal cultures by co-culturing with astrocytes and oligodendrocyte precursor cells (complex culture). We evaluated and compared neuronal activity (e.g., network formation and maturation), cellular composition in long-term culture, and the transcriptome of the two cultures. Compared to simple cultures, neurons from complex co-cultures exhibited earlier synapse and network development and maturation, which was supported by localized synaptophysin expression, up-regulation of genes involved in mature neuronal processes, and synchronized neural network activity. Also, mature oligodendrocytes and reactive astrocytes were only detected in complex cultures upon transcriptomic analysis of age-matched cultures. Functionally, the GABA antagonist bicuculline had a greater influence on bursting activity in complex versus simple cultures. Collectively, the cellular complexity of brain-on-a-chip systems intrinsically develops cell type-specific phenotypes relevant to the brain while accelerating the maturation of neuronal networks, important features underdeveloped in traditional cultures.
脑芯片系统旨在使用传统的工程平台上的体外细胞培养来模拟大脑活动。它是一种非侵入性的工具，用于筛选新药、评估毒物和阐明疾病机制。然而，这些系统对大脑功能的成功再现依赖于细胞培养的复杂性。在这项研究中，我们通过与星形胶质细胞和少突胶质细胞前体细胞（复杂培养）共培养来增加传统（简单）神经元培养的细胞复杂性。我们评估和比较了两种培养物的神经元活性（例如，网络形成和成熟）、长期培养中的细胞组成以及转录组。与简单培养相比，来自复杂共培养的神经元表现出更早的突触和网络发育和成熟，这得到了局部突触素表达、成熟神经元过程中涉及的基因上调以及同步神经网络活动的支持。此外，只有在对年龄匹配的培养物进行转录组分析时，才会在复杂培养物中检测到成熟的少突胶质细胞和反应性星形胶质细胞。在功能上，GABA 拮抗剂荷包牡丹碱对复杂培养物中的爆发活动的影响大于对简单培养物的影响。总的来说，脑芯片系统的细胞复杂性内在地发展出与大脑相关的细胞特异性表型，同时加速神经元网络的成熟，这是传统培养物中尚未充分发展的重要特征。
