Introduction The effects of anti-cancer drugs on 3D spheroids can be assessed using electrical impedance spectroscopy (EIS) per-formed on microelectrode arrays (MEAs). However, in planar MEAs, the solid substrate limits nutrient diffusion to the bottom of the spheroids. In addition, spheroid placement and stabilization on planar MEAs are challenging. To overcome these limitations, mesh MEAs are investigated here, allowing the culture medium to surround 3D cell-cultured spheroids and improving mass transport conditions. Methods The mesh MEA (MultiChannelSystem MCS GmbH, Germany) comprised of 60 electrodes of 30 µm diameter and an inter-electrode spacing of 200 µm, fabricated on a polyimide mesh. Spheroids of 250 µm size were generated from the HT-29 cell line and cultivated on mesh MEAs for 96 hours. Navitoclax (10 µM) was tested against untreated sphe-roids (n = 5 per group). EIS was performed using the impedance analyzer ISX-3 (Sciospec GmbH, Germany). The rel-ative impedance change (RIC) was calculated to assess mesh MEA performance. Results The mesh MEA enabled easy placement and stable holding of spheroids. Cells proliferated around the mesh structure, allowing for impedance measurements inside the spheroid and at its periphery. In both control and Navitoclax-treated groups, RIC increased up to 48 hours. After this time point, RIC values stabilized in the control group, whereas a re-duction in RIC was observed in the Navitoclax-treated spheroids. On rigid planar MEAs, a large percentage of sphe-roids moved during culture. Additionally, impedance signals were declining, potentially due to the formation of a ne-crotic core within the spheroids. Conclusion Mesh MEAs provide a robust platform for stable spheroid cultivation and long-term EIS measurements. Drug-induced effects can be reliably detected using mesh MEAs for 3D in vitro drug screening applications. Future work will focus on testing drug effects on spheroids derived from different cell lines on the same mesh MEA.