MDCKII cells were seeded inside the scaffold devices and incubated to grow for 3 days. After stabilization of the devices for several minutes, we recorded the electrical signal (transient current response) to investigate the effects of EGTA addition. Specifically, a 100 mM solution of EGTA was diluted in cell medium (Alfa Aesar EGTA 0.5 M aqueous solution, J60767) and injected into the scaffold through the fluidic circuit in a steady manner. The transistor recording was carried out for several minutes after EGTA injection. The following parameters were used during the transient response measurements: VDS = −0.3 V, VGS = 0.3 V, on time t = 15 s. Data analysis was performed to fit the time constant for each individual pulse (13). Control experiments were carried out by exposing a cell-free scaffold to the EGTA solution for a certain period of time, as shown in the normalized data of fig. S8C.

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