Abstract
Background:
The cardiac autonomic nervous system maintains the cardiac homeostasis and alterations of it can be involved in cardiac disorders. However, these mechanisms remain poorly understood due to the lack of proper human cell models.
Methods:
A neurocardiac co-culture model was created using exclusively cells derived from human induced pluripotent stem cells (iPSCs) in a 2 chambers silicon insert.
Findings:
iPSC-cardiomyocytes (CMs) and iPSC-sympathetic neurons (SNs) were co-cultured in two separate chambers and, after insert removal, iPSC-SNs formed axons projecting towards the CMs, and visualized with immunostaining. The beat rate of iPSC-CMs was measured using the MEA system and was stable after 7 days of co-culture. A significant increase in the beat rate of iPSC-CMs was observed after nicotine treatment that had no effect on iPSC-CMs in monoculture. On the contrary, after treatment with α-bungarotoxin, which bind to nicotinic receptors blocking neural transmission, the beat rate of iPSC-CMs in co-culture was unaffected. iPSC-derived CMs responded positively to isoproterenol treatment with a significant increase of the beat rate and the subsequent propranolol administration decreased the beat activity of the cells. After 7 days of co-culture a significant decrease of the number of vesicles stained with FFN270 was observed in nicotine treated cells in comparison to the co-culture at baseline resulting in an effective release of the neurotransmitters, showing the functional exocytosis process.
Interpretation:
The proposed neurocardiac model recapitulates the neuro-cardiac axis and provides a promising modelling tool for the study of molecular mechanisms involved in the in health and disease.
Funding: ITAT1047