Advanced in-vitro tools have emerged to address ethical and environmental concerns in bio-related research¹. Traditional methods maintain in-vitro cultures in incubators with manual media changes and monitoring. We developed a perfusion circuit for nearly intervention-free long-term neural cell culture maintenance and electrophysiological measurement. It incorporates an adaptable media supply system, a gas-exchange unit, a temperature controller, a filtering system and cell-hosting compartment. This device, integrated with a reader-stimulator for electrophysiology and neuromodulation, permits significant insights into synaptic plasticity, essential for learning and memory², along with detailed tracking of cell-culture maturation.

The automated closed-loop perfusion circuit is driven by multiple peristaltic pumps to ensure systematic media refreshment, enabling us to correlate neuronal activity with the consumption rate of media components. Additionally, the multiple pumps allow differential pressures to optimize a filtration system designed to reuse cleaned media for extended periods. The counter-flow filtration system, used in the perfusion circuit, provides an effective mechanism for neuronal media filtration within the perfusion circuit throughout the experiment. These features make the perfusion circuit ideal for neuroscientific research, disease modelling, and drug testing.

Neural cultures were grown on glass planar multielectrode arrays and integrated into the perfusion circuit³. Spent media and measured electrophysiological activity was measured over time. Our results confirmed that the perfusion circuit supports neural cell maintenance for over six months. With further development, we believe the perfusion circuit will become a valuable in-vitro tool, enhancing sustainability in neuronal studies and reducing animal testing in early-stage neuronal disease modelling and drug testing.