A Three-Dimensional Arrayed Microfluidic Blood–Brain Barrier Model With Integrated Electrical Sensor Array

<italic>Objective:</italic> The blood–brain barrier (BBB) poses a unique challenge to the development of therapeutics against neurological disorders due to its impermeabi-lity to most of the chemical compounds. Most <italic> in vitro</italic> BBB models have limitations in mimicking <italic>in vivo</italic> conditions and functions. Here, we show a co-culture microfluidic BBB-on-a-chip that provides interactions between neurovascular endothelial cells and neuronal cells across a porous polycarbonate membrane, which better mimics the <italic>in vivo</italic> conditions, as well as allows <italic>in vivo</italic> level shear stress to be applied. <italic>Methods:</italic> A 4 × 4 intersecting microchannel array forms 16 BBB sites on a chip, with a multielectrode array integrated to measure the transendothelial electrical resistance (TEER) from all 16 different sites, which allows label-free real-time analysis of the barrier function. Primary mouse endothelial cells and primary astrocytes were co-cultured in the chip while applying <italic>in vivo</italic> level shear stress. The chip allows the barrier function to be analyzed through TEER measurement, dextran permeability, as well as immunostaining. <italic>Results:</italic> Co-culture between astrocytes and endothelial cells, as well as <italic>in vivo</italic> level shear stress applied, led to the formation of tighter junctions and significantly lower barrier permeability. Moreover, drug testing with histamine showed increased permeability when using only endothelial cells compared to almost no change when using co-culture. <italic>Conclusion: </italic> Results show that the developed BBB chip more closely mimics the <italic>in vivo</italic> BBB environment. <italic>Significance:</italic> The developed multisite BBB chip is expected to be used for screening drug by more accurately predicting their permeability through BBB as well as their toxicity.