Generation of parasympathetic neurons from hiPSC that reproduce the electrophysiological properties of native neurons and modulate the activity of hiPSC-atrial cardiomyocytes
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Peripheral parasympathetic ganglia lie adjacent to their target organs; their dysfunction contributes to diseases like atrial fibrillation. Due to their challenging accessibility for primary culture, we provide a step-by-step protocol to generate parasympathetic neurons from human induced pluripotent stem cells (hiPSC). They reproduce numerous native physiological features: cholinergic and autonomic markers, functional nicotinic receptors and electrophysiological properties. We also describe a co-culture system with hiPSC-atrial cardiomyocytes allowing neuromodulation of cardiomyocyte activity and offering a platform for drug discovery and disease modelling.
Highlights
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Steps to generate parasympathetic neurons from human induced pluripotent stem cells
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Guidance of the functional assessment of the neurons (whole-cell patch clamp and Ca 2+ imaging)
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Instructions on how to co-culture these neurons with hiPSC-derived atrial cardiomyocytes
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Guidance of the functional assessment of the neuronal modulation of cardiomyocyte activity
