Abstract
The cardiac autonomic nervous system (CANS) is widely recognized as an important player in many cardiac disorders, such as arrhythmogenic cardiomyopathy, catecholaminergic polymorphic ventricular tachycardia, ventricular tachyarrhythmia and heart failure [1]. The mechanisms of neuronal control of cardiac disease remain elusive, largely due to the lack of proper human cell models. To overcome this limitation, we have created an in vitro neurocardiac model system by using a silicon-based two-wells insert (Ibidi) with a defined cell-free gap for the co-culture of human iPSC-derived cardiomyocytes (iPSC-CMs) and sympathetic neurons (iPSC-SNs). Here we describe the assessment of the physiological activity of this co-culture model, using a Maestro Edge Multi-Electrode Array (MEA, Axion Biosystems) (see Figure 1). In particular, by evaluating the 3D surface reconstruction of the expression of the presynaptic marker protein Synapsin 1 (Syn1) and the sarcomeric protein a-actinin, we observed a punctate staining of varicosities suggesting multiple synaptic connections between the iPSC-CMs and the iPSC-SNs. In addition, to verify whether the iPSC-SNs were forming functional connections with the iPSC-CMs and therefore affect their activity, we have evaluated the electrical properties of iPSC-CMs after nicotine treatment. Preliminary evidence indicates that in the presence of iPSC-SNs the beating rate of iPSC-CMs is selectively increased after nicotine stimulation. Furthermore, it has been observed that through electrical stimulation of iPSC-SNs it is also possible to modulate the beating rate of iPSC-CMs. This neurocardiac model provides a promising tool to model a wide range of cardiac but also neurological pathologies, as well as for drug screening and personalized approaches.
Funding acknowledgements:
Funded by the European Regional Development Fund and Interreg V-A Italy-Austria 2014-2020 and by the Department of Innovation, Research and University of the Autonomous Province of Bolzano-South Tyrol (Italy).