Overflow Microfluidic Networks: Application to the Biochemical Analysis of Brain Cell Interactions in Complex Neuroinflammatory Scenarios
Overflow Microfluidic Networks: Application to the Biochemical Analysis of Brain Cell Interactions in Complex Neuroinflammatory Scenarios
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DOI：
10.1021/ac302094z
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发表时间：
2012-11-20
影响因子：
7.4
通讯作者：
Matteoli, Michela
中科院分区：
文献类型：
--
作者：
Bianco, Fabio;Tonna, Noemi;Matteoli, Michela
神经炎症在神经退行性疾病中起着核心作用，涉及不同脑细胞类型之间的大量相互作用。揭示神经炎症中细胞-细胞相互作用的复杂性对于阐明所涉及的分子机制和提高药物开发的功效至关重要。在这里，我们提供了一种通用的分析方法，专门解决细胞间的通信，使用原代脑细胞，微流体装置，和多参数读出方法。将不同的细胞类型接种在微流体网络的单独的室中，使得培养条件可以独立地控制，并且可以用不同的刺激物选择性地引发单细胞类型。当腔室被微流体连接时，可以通过分析形态学、活力、钙动力学和电生理学参数来精细地监测每种细胞类型的具体贡献。我们通过研究来自两个不同脑区（皮质和海马）的星形胶质细胞在两种类型的神经炎性损伤（即代谢应激和暴露于淀粉样蛋白β原纤维）中对神经元活力的作用，证实了这种方法，并证明了神经元生理病理学的胶质细胞控制的区域差异。特别是，我们表明，在代谢应激期间，皮质而不是海马星形胶质细胞发挥神经保护作用;此外，在暴露于A β + IL-1 β的炎症加剧的情况下，海马而不是皮质星形胶质细胞对神经元发挥有害作用。除了对神经胶质在神经炎症中的作用提出新的见解外，本文提出的方法还代表了一种有前途的工具，用于解决各种生物和生化现象，其特征在于多种细胞类型的复杂相互作用。
Neuroinflammation plays a central role in neurodegenerative diseases and involves a large number of interactions between different brain cell types. Unraveling the complexity of cell-cell interaction in neuroinflammation is crucial for both clarifying the molecular mechanisms involved and increasing efficacy in drug development. Here, we provide a versatile analytical method for specifically addressing cell-to-cell communication, using primary brain cells, a microfluidic device, and a multiparametric readout approach. Different cell types are plated in separate chambers of a microfluidic network so that culturing conditions can be independently controlled and single cell types can be selectively primed with different stimuli. When chambers are microfluidically connected, the specific contribution of each cell type can be finely monitored by analyzing morphology, vitality, calcium dynamics, and electrophysiology parameters. We exemplify this approach by examining the role of astrocytes derived from two different brain regions (cortex and hippocampus) on neuronal viability in two types of neuroinflammatory insults, namely, metabolic stress and exposure to amyloid beta fibrils, and demonstrate regional differences in glial control of neuronal physiopathology. In particular, we show that during metabolic stress, cortical but not hippocampal astrocytes play a neuroprotective role; also, in an exacerbated inflammatory scenario consisting in the exposure to A beta + IL-1 beta, hippocampal but not cortical astrocytes play a detrimental role on neurons. Aside from bringing novel insights into the glial role in neuroinflammation, the method presented here represents a promising tool for addressing a wide range of biological and biochemical phenomena, characterized by a complex interaction of multiple cell types.