Department of Chemical, Biological, and Bioengineering, North Carolina Agricultural and Technical State University, Greensboro, NC, USA.
Methods Mol Biol. 2022;2492:117-128. doi: 10.1007/978-1-0716-2289-6_6.
Brain-on-a-chip is a miniaturized engineering platform to mimic the structural and functional aspects of brain tissue. We describe a method to construct a three-dimensional (3D) brain-on-a-chip in this chapter. We firstly portray the method of a brain-on-a-chip model with cocultured mice neurons, microglia, and astrocytes to mimic brain tissue and membrane-free perfusion with endothelial cells, in which we successfully build the blood-brain barrier to screen neurotoxicity. Then we describe a method to construct a brain-on-a-chip with human induced pluripotent stem cell (iPSC)-derived neurons and astrocytes to simulate human brain behavior. This platform consists of neuronal tissue with extracellular matrix (ECM)-embedded GABAergic neurons and astrocytes and a perfusion channel with dynamic flow. We also include the broader applicability test of this model using an organophosphate (OP), malathion, to induce acute and chronic neurotoxicity, and then using butyrylcholinesterase (BuChE) as an exogenous bioscavenger of OP. Following the methods listed in this chapter, we are able to measure the neurotoxic effects on construct integrity, viability, and total AChE and BuChE activity.
脑芯片是一种微型工程平台，用于模拟脑组织的结构和功能方面。在本章中，我们将描述构建三维（3D）脑芯片的方法。我们首先描述了一种使用共培养的小鼠神经元、小胶质细胞和星形胶质细胞模拟脑组织的脑芯片模型的方法，并采用无膜灌流的方法模拟内皮细胞，成功构建了血脑屏障以筛选神经毒性。然后，我们描述了一种使用人诱导多能干细胞（iPSC）衍生的神经元和星形胶质细胞构建脑芯片的方法，以模拟人类大脑的行为。该平台由神经元组织和具有动态流动的灌注通道组成，其中神经元组织嵌入 GABA 能神经元和星形胶质细胞的细胞外基质（ECM）。我们还通过使用有机磷（OP）马拉硫磷来测试该模型的更广泛适用性，以诱导急性和慢性神经毒性，然后使用丁酰胆碱酯酶（BuChE）作为 OP 的外源性生物清除剂。按照本章列出的方法，我们能够测量对构建体完整性、活力以及总 AChE 和 BuChE 活性的神经毒性影响。
