title: Development and function of human cerebral cortex neural networks from pluripotent stem cells in vitro.
abstract: A key aspect of nervous system development, including that of the cerebral cortex, is the formation of higher-order neural networks. Developing neural networks undergo several phases with distinct activity patterns in vivo, which are thought to prune and fine-tune network connectivity. We report here that human pluripotent stem cell (hPSC)-derived cerebral cortex neurons form large-scale networks that reflect those found in the developing cerebral cortex in vivo. Synchronised oscillatory networks develop in a highly stereotyped pattern over several weeks in culture. An initial phase of increasing frequency of oscillations is followed by a phase of decreasing frequency, before giving rise to non-synchronous, ordered activity patterns. hPSC-derived cortical neural networks are excitatory, driven by activation of AMPA- and NMDA-type glutamate receptors, and can undergo NMDA-receptor-mediated plasticity. Investigating single neuron connectivity within PSC-derived cultures, using rabies-based trans-synaptic tracing, we found two broad classes of neuronal connectivity: most neurons have small numbers (<10) of presynaptic inputs, whereas a small set of hub-like neurons have large numbers of synaptic connections (>40). These data demonstrate that the formation of hPSC-derived cortical networks mimics in vivo cortical network development and function, demonstrating the utility of in vitro systems for mechanistic studies of human forebrain neural network biology.
authors.affiliation: Wellcome Trust/CRUK Gurdon Institute and Department of Biochemistry, University of Cambridge, Tennis Court Road, Cambridge CB2 1QN, UK.
authors.forename: Peter
authors.initials: P
authors.lastname: Kirwan
authors.affiliation: Wellcome Trust/CRUK Gurdon Institute and Department of Biochemistry, University of Cambridge, Tennis Court Road, Cambridge CB2 1QN, UK.
authors.forename: Benita
authors.initials: B
authors.lastname: Turner-Bridger
authors.affiliation: Wellcome Trust/CRUK Gurdon Institute and Department of Biochemistry, University of Cambridge, Tennis Court Road, Cambridge CB2 1QN, UK.
authors.forename: Manuel
authors.initials: M
authors.lastname: Peter
authors.affiliation: Wellcome Trust/CRUK Gurdon Institute and Department of Biochemistry, University of Cambridge, Tennis Court Road, Cambridge CB2 1QN, UK.
authors.forename: Ayiba
authors.initials: A
authors.lastname: Momoh
authors.affiliation: Department of Physiology, Development and Neuroscience, University of Cambridge, Tennis Court Road, Cambridge CB2 1QN, UK.
authors.forename: Devika
authors.initials: D
authors.lastname: Arambepola
authors.affiliation: Department of Physiology, Development and Neuroscience, University of Cambridge, Tennis Court Road, Cambridge CB2 1QN, UK.
authors.forename: Hugh P C
authors.initials: HP
authors.lastname: Robinson
authors.affiliation: Wellcome Trust/CRUK Gurdon Institute and Department of Biochemistry, University of Cambridge, Tennis Court Road, Cambridge CB2 1QN, UK rick@gurdon.cam.ac.uk.
authors.forename: Frederick J
authors.initials: FJ
authors.lastname: Livesey
mesh_terms: D002540:Cerebral Cortex / Q000166:cytology / Q000254:growth & development*; D049229:Dendritic Spines / Q000648:ultrastructure; D006801:Humans; D007091:Image Processing, Computer-Assisted; D066298:In Vitro Techniques / Q000379:methods*; D008856:Microscopy, Fluorescence; D009415:Nerve Net / Q000166:cytology / Q000254:growth & development*; D009473:Neuronal Plasticity / Q000502:physiology; D018408:Patch-Clamp Techniques; D039904:Pluripotent Stem Cells / Q000502:physiology*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D059010:Single-Cell Analysis; D014741:Video Recording
keywords: Cerebral cortex; Human; Networks; Neural development; Stem cells