神经炎症在神经系统疾病中根据风险因素和神经胶质细胞相互作用会产生有益或有害的影响。然而，由于细胞间相互作用的复杂性和缺乏具有生理病理相关性的神经炎症模型，神经炎症的研究一直具有挑战性。在这里，我们描述了我们的三维微流控多细胞人类神经培养模型，被称为“芯片上的大脑”（BoC）。它以受控的方式阐明了神经胶质细胞之间的相互作用，并复现了主要神经系统疾病的病理特征：痴呆、脑肿瘤和脑水肿。该平台包括一个趋化模块，相比其他趋化模型的几个小时，它提供了一个持续一周的稳定趋化梯度。此外，与用混合型微胶质细胞培养的传统脑模型相比，我们的BoC能够将与疾病相关的微胶质细胞从异质种群中分离出来，并在三维空间中实现选择性的神经胶质细胞相互作用。这样一来，我们在多种疾病的BoC模型中验证了活跃的先天免疫细胞是导致突触损伤和神经元丧失的风险因素，这有助于解释神经胶质细胞的相互作用。该方案描述了如何制备和应用我们的人体BoC，在实时操作和终点分析中进行。设置装置和细胞制备需要2天，以疾病条件下发展脑模型需要1-9周，并且进行分析需要2-3天。该方案要求具备基本分子生物学技术的研究人员进行至少1个月的培训。总的来说，我们的人体BoCs是可靠而宝贵的平台，可用于研究涉及神经炎症的病理机制，并评估调节神经炎症的治疗策略在神经系统疾病中的效果。© 2023. Springer Nature Limited.

Neuroinflammation has either beneficial or detrimental effects, depending on risk factors and neuron-glia interactions in neurological disorders. However, studying neuroinflammation has been challenging due to the complexity of cell-cell interactions and lack of physio-pathologically relevant neuroinflammatory models. Here, we describe our three-dimensional microfluidic multicellular human neural culture model, referred to as a 'brain-on-a-chip' (BoC). This elucidates neuron-glia interactions in a controlled manner and recapitulates pathological signatures of the major neurological disorders: dementia, brain tumor and brain edema. This platform includes a chemotaxis module offering a week-long, stable chemo-gradient compared with the few hours in other chemotaxis models. Additionally, compared with conventional brain models cultured with mixed phenotypes of microglia, our BoC can separate the disease-associated microglia out of heterogeneous population and allow selective neuro-glial engagement in three dimensions. This provides benefits of interpreting the neuro-glia interactions while revealing that the prominent activation of innate immune cells is the risk factor leading to synaptic impairment and neuronal loss, validated in our BoC models of disorders. This protocol describes how to fabricate and implement our human BoC, manipulate in real time and perform end-point analyses. It takes 2 d to set up the device and cell preparations, 1-9 weeks to develop brain models under disease conditions and 2-3 d to carry out analyses. This protocol requires at least 1 month training for researchers with basic molecular biology techniques. Taken together, our human BoCs serve as reliable and valuable platforms to investigate pathological mechanisms involving neuroinflammation and to assess therapeutic strategies modulating neuroinflammation in neurological disorders.© 2023. Springer Nature Limited.