Human heart tissues grown as three-dimensional spheroids and consisting of different cardiac cell types derived from pluripotent stem cells (hiPSCs) recapitulate aspects of human physiology better than standard two-dimensional models in vitro. They typically consist of less than 5000 cells and are used to measure contraction kinetics although not contraction force. By contrast, engineered heart tissues (EHTs) formed around two flexible pillars, can measure contraction force but conventional EHTs often require between 0.5 and 2 million cells. This makes large-scale screening of many EHTs costly. Our goals here were (i) to create a physiologically relevant model that required fewer cells than standard EHTs making them less expensive, and (ii) to ensure that this miniaturized model retained correct functionality. We demonstrated that fully functional EHTs could be generated from physiologically relevant combinations of hiPSC-derived cardiomyocytes (70%), cardiac fibroblasts (15%) and cardiac endothelial cells (15%), using as few as 1.6 × 10⁴ cells. Our results showed that these EHTs were viable and functional up to 14 days after formation. The EHTs could be electrically paced in the frequency range between 0.6 and 3 Hz, with the optimum between 0.6 and 2 Hz. This was consistent across three downscaled EHT sizes tested. These findings suggest that miniaturized EHTs could represent a cost-effective microphysiological system for disease modelling and examining drug responses particularly in secondary screens for drug discovery.

Microelectrode arrays (MEAs) are widely used in biological application to locally stimulate and record the electrical activity of living cells. Here, a novel fabrication process for a carbon nanotube (CNT)-based MEA integrated on the backside of a free standing stretchable membrane is reported. The new process flow overcomes the manually intensive procedures used in the previous works. The microfabricated upside-down CNT MEA consists of microelectrodes with an area of 110 μm2 covered with cobalt-grown CNTs. The surface area enhancement and the foamlike morphology of the CNTs allow an increase of the charge injection per unit area at the electrode–electrolyte interface, resulting in a significantly lower electrochemical impedance of the electrodes. In particular, at 1 kHz, the fabricated CNT-MEA electrodes show a reduction of the overall impedance up to 96% in comparison with benchmark TiN electrodes. The obtained results confirm the effectiveness of the proposed surface texturing through CNT integration. Moreover, the quality and the morphology as well as the biocompatibility of the fabricated CNT-based electrodes were assessed. The obtained results demonstrate that significant improvement can be achieved by integrating structured nanoporous material on MEAs.