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

  1. Alison M. Thomas
  2. Isabella Noelle Chiong
  3. Joana F Neves
  4. Andrew Tinker
  5. Franziska Denk
  6. Laura Fedele
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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.

Highlights

  • Steps to generate parasympathetic neurons from human induced pluripotent stem cells

  • Guidance of the functional assessment of the neurons (whole-cell patch clamp and Ca 2+ imaging)

  • Instructions on how to co-culture these neurons with hiPSC-derived atrial cardiomyocytes

  • Guidance of the functional assessment of the neuronal modulation of cardiomyocyte activity

Graphical abstract

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