Multi-level characterization of balanced inhibitory-excitatory cortical neuron network derived from human pluripotent stem cells

PLoS One. 2017 Jun 6;12(6):e0178533. doi: 10.1371/journal.pone.0178533. eCollection 2017.

Abstract

Generation of neuronal cultures from induced pluripotent stem cells (hiPSCs) serve the studies of human brain disorders. However we lack neuronal networks with balanced excitatory-inhibitory activities, which are suitable for single cell analysis. We generated low-density networks of hPSC-derived GABAergic and glutamatergic cortical neurons. We used two different co-culture models with astrocytes. We show that these cultures have balanced excitatory-inhibitory synaptic identities using confocal microscopy, electrophysiological recordings, calcium imaging and mRNA analysis. These simple and robust protocols offer the opportunity for single-cell to multi-level analysis of patient hiPSC-derived cortical excitatory-inhibitory networks; thereby creating advanced tools to study disease mechanisms underlying neurodevelopmental disorders.

MeSH terms

  • Astrocytes / cytology
  • Astrocytes / physiology*
  • Cell Differentiation / physiology
  • Cells, Cultured
  • Coculture Techniques*
  • Electrophysiological Phenomena
  • GABAergic Neurons / cytology
  • GABAergic Neurons / physiology*
  • Humans
  • Induced Pluripotent Stem Cells / cytology
  • Induced Pluripotent Stem Cells / physiology*
  • Single-Cell Analysis

Grants and funding

This study was supported and funded by Neuroscience Campus Amsterdam and EU MSCA-ITN CognitionNet (FP7-PEOPLE-2013-ITN 607508).