In recent decades, three-dimensional (3D) cancer cell models have attracted increasing interest in the field of drug screening due to their significant advantages in more accurate simulations of heterogeneous tumor behavior in vivo compared to two-dimensional models. Furthermore, drug sensitivity testing based on 3D cancer cell models can provide more reliable in vivo efficacy prediction. The gold standard fluorescence staining is hard to achieve real-time and label-free viability monitoring in 3D cancer cell models. In this study, a microgroove impedance sensor (MGIS) was specially developed for the dynamic and noninvasive monitoring of 3D cell viability. 3D cancer cells were trapped in microgrooves with gold electrodes on opposite walls for in situ impedance measurement. The change in the number of live cells caused inversely proportional changes to the impedance magnitude of the entire cell/Matrigel construct and reflected the proliferation and apoptosis of the 3D cells. It was confirmed that the 3D cell viability detected by the MGIS was highly consistent with the standard live/dead staining by confocal microscope characterization. Furthermore, the accuracy of the MGIS was validated quantitatively using a 3D lung cancer model and sophisticated drug sensitivity testing. In addition, the parameters of the MGIS in the measurement experiments were optimized in detail using simulations and experimental validation. The results demonstrated that the MGIS coupled with 3D cell culture would be a promising platform to improve the efficiency and accuracy of cell-based anticancer drug screening in vitro.

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