Functional Coupling with Cardiac Muscle Promotes Maturation of hPSC-Derived Sympathetic Neurons

Cell Stem Cell. 2016 Jul 7;19(1):95-106. doi: 10.1016/j.stem.2016.05.002. Epub 2016 Jun 16.


Neurons derived from human pluripotent stem cells (hPSCs) are powerful tools for studying human neural development and diseases. Robust functional coupling of hPSC-derived neurons with target tissues in vitro is essential for modeling intercellular physiology in a dish and to further translational studies, but it has proven difficult to achieve. Here, we derive sympathetic neurons from hPSCs and show that they can form physical and functional connections with cardiac muscle cells. Using multiple hPSC reporter lines, we recapitulated human autonomic neuron development in vitro and successfully isolated PHOX2B::eGFP+ neurons that exhibit sympathetic marker expression and electrophysiological properties and norepinephrine secretion. Upon pharmacologic and optogenetic manipulation, PHOX2B::eGFP+ neurons controlled beating rates of cardiomyocytes, and the physical interactions between these cells increased neuronal maturation. This study provides a foundation for human sympathetic neuron specification and for hPSC-based neuronal control of organs in a dish.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Animals, Newborn
  • Basic Helix-Loop-Helix Transcription Factors / metabolism
  • Cell Differentiation* / genetics
  • Cell Line
  • Flow Cytometry
  • Gene Expression Regulation, Developmental
  • Genes, Reporter
  • Green Fluorescent Proteins / metabolism
  • Heart Ventricles / cytology
  • Hedgehog Proteins / metabolism
  • Homeodomain Proteins / metabolism
  • Humans
  • Mice, Inbred C57BL
  • Myocytes, Cardiac / cytology*
  • Myocytes, Cardiac / metabolism
  • Neurons / cytology*
  • Neurons / metabolism
  • Optogenetics
  • Phenotype
  • Pluripotent Stem Cells / cytology*
  • Pluripotent Stem Cells / metabolism
  • Sympathetic Nervous System / cytology*
  • Transcription Factors / metabolism
  • Wnt Signaling Pathway / genetics


  • ASCL1 protein, human
  • Basic Helix-Loop-Helix Transcription Factors
  • Hedgehog Proteins
  • Homeodomain Proteins
  • NBPhox protein
  • SHH protein, human
  • Transcription Factors
  • enhanced green fluorescent protein
  • Green Fluorescent Proteins