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bioRxiv · 10.1101/2025.10.23.684157

Generation of parasympathetic neurons from hiPSC that reproduce the electrophysiological properties of native neurons and modulate the activity of hiPSC-atrial cardiomyocytes

Thomas, A. M.·Chiong, I. N.·Neves, J. F.·Tinker, A.·Denk, F.·Fedele, L.
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Abstract

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. HighlightsO_LISteps to generate parasympathetic neurons from human induced pluripotent stem cells C_LIO_LIGuidance of the functional assessment of the neurons (whole-cell patch clamp and Ca2+ imaging) C_LIO_LIInstructions on how to co-culture these neurons with hiPSC-derived atrial cardiomyocytes C_LIO_LIGuidance of the functional assessment of the neuronal modulation of cardiomyocyte activity C_LI Graphical abstract O_FIG O_LINKSMALLFIG WIDTH=198 HEIGHT=200 SRC="FIGDIR/small/684157v1_ufig1.gif" ALT="Figure 1"> View larger version (44K): org.highwire.dtl.DTLVardef@1e77cfborg.highwire.dtl.DTLVardef@be6cbforg.highwire.dtl.DTLVardef@1d58aa6org.highwire.dtl.DTLVardef@81ba0e_HPS_FORMAT_FIGEXP M_FIG C_FIG

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Thomas, A. M., Chiong, I. N., Neves, J. F., Tinker, A., Denk, F., Fedele, L.. 2025-10-24. Generation of parasympathetic neurons from hiPSC that reproduce the electrophysiological properties of native neurons and modulate the activity of hiPSC-atrial cardiomyocytes. https://doi.org/10.1101/2025.10.23.684157

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