A novel three-dimensional brain-on-chip model using human iPSC-derived GABAergic neurons and astrocytes

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-chip in this chapter. We firstly portray the method of a brain-on-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 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.