Human cardiac innervation-on-a-chip platform recapitulates neurocardiac interactions and supports future disease modeling

Abstract

Abstract The cardiac autonomic nervous system regulates cardiac function through innervation, and dysfunction of the neuronal and cardiovascular crosstalk has been linked to various pathologies. Here, we present a cardiac innervation-on-a-chip model by combining human induced pluripotent stem cell (hiPSC)-derived cortical neurons, postganglionic sympathetic neurons, and ventricular cardiomyocytes in a compartmentalized microfluidic device called a 3D3C chip to generate physiologically relevant in vitro brain-heart axis model. The 3D3C chip allowed successful multiculturing of the cell types in distinct compartments and formation of axonal connections between them. Using integrated microelectrode arrays (MEA), we demonstrated the progressive development of spontaneous electrophysiological activity in both neurons and cardiomyocytes over time. This subsequently allowed for the pharmacological stimulation of neuronal activity followed by axon-mediated cardiac responses and increased neurotransmitter release. Moreover, innervated cardiomyocytes exhibited both enhanced structural maturation and electrophysiological functionality in the multiculture. This advanced cardiac innervation-on-a-chip provides a powerful in vitro platform for examining disease-related mechanisms of the brain–heart axis.