Cardiotoxicity is one of the main side effects limiting development of new medicines and resulting in withdrawal of drugs from the market. There are different mechanisms of drug-induced cardiotoxicity, including arrhythmia, disruption of mitochondria function, apoptosis, altered growth factor signalling [1]. Heart on a chip model for drug testing seems attractive approach to evaluate the possible cardiotoxic effects on cell biological, but mostly electrophysiological properties. Our aim was to create a 3D system with a biocompatible scaffold, coated with a conductive layer, suitable for culturing and maturation of human induced pluripotent stem cell derived cardiomyocytes (hiPSC-CMs) and compatible with electrophysiology parameter measurement techniques. Methods Elastic scaffold was fabricated by electrospinning of gelatin-glucose microfibers followed by the vapor-phase and electrochemical deposition of conductive polymer polypyrrole (PPy) to create electroconductive layer. hiPSC-CMs at different maturation levels (at 6 vs 14 weeks) were used to test biocompatibility of the scaffold. To increase cell attachment, scaffold was treated with 0.1 M HCl and coated with Geltrex. Cells were stained with fluorescent dyes (Calcein AM, DAPI). Electrophysiological properties were analysed using intracellular calcium imaging (Cal520) and action potential recordings with sharp electrodes. HiPSC-CMs on coverslips were used as a 2D control. Results Calcein AM staining showed that high number of hiPSC-CMs attached to gelatine/glucose/PPy scaffold proving that the scaffold is biocompatible and cells remain viable even after long term incubation (> 2 weeks). HiPSC-CMs at lower maturity (6 weeks) had better attachment properties to it, indicating that more mature cardiomyocytes (14 weeks) gradually lose their capacity to attach to PPy surface. Intracellular calcium imaging showed that both types of cells generated synchronous spontaneous calcium oscillations on gelatine/glucose/PPy scaffolds. Action potential recordings in 3D were similar to those in 2D, and confirmed differences in hiPSC-CMs maturation levels – at lower maturation level hiPSC-CMs had lower upstroke velocity as compared to more matured hiPSC-CMs. Conclusions Gelatine/glucose/PPy scaffold is biocompatible for hiPSC-CMs cultivation, and suitable for evaluation of cardiomyocyte electrophysiological properties. Such electromechanoactive scaffold can be further employed for development of heart-on-a-chip model.