On-chip real-time impedance monitoring of hiPSC-derived and artificial basement membrane-supported endothelium

Recent advances have shown the high sensibility of electrochemical impedance spectroscopy in real-time monitoring of cell barriers on a chip. Here, we applied this method to the investigation of human induced pluripotent stem cell (hiPSC) derived and artificial basement membrane (ABM) supported endothelial barrier. The ABM was obtained by self-assembling type IV collagen and laminin with a monolayer of crosslinked gelatin nanofibers. The hiPSCs were differentiated into brain microvascular endothelial cells (BMECs) and then plated on the ABM. After incubation for two days, the ABM-BMEC assembly was placed as a tissue insert into a microfluidic device for culture and real-time impedance monitoring over days. We found a significantly enhanced stability of the BMEC barrier in a serum-free and bromodeoxyuridine (BrdU) containing culture medium compared to the conventional culture due to the restricted cell proliferation. We also found that the BMEC barrier was sensitive to stimuli such as thrombin and that the change of the barrier impedance was mainly due to the change of the cell layer resistance. We can thus advocate this method to investigate the integrity of the cell barrier and the barrier-based assays.