Cell Stem Cell
Volume 22, Issue 5, 3 May 2018, Pages 684-697.e9
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Article
Efficient Generation of CA3 Neurons from Human Pluripotent Stem Cells Enables Modeling of Hippocampal Connectivity In Vitro

https://doi.org/10.1016/j.stem.2018.04.009Get rights and content
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Highlights

  • Differentiation of human CA3 pyramidal neuron is modeled using ESCs/iPSCs

  • RNA-seq and immunocytochemistry revealed pyramidal neuron diversity

  • Rabies virus tracing showed a connection between ESC-derived DG and CA3 neurons

  • Schizophrenia hiPSC-derived DG-CA3 co-cultures present deficits in hippocampal activity

Summary

Despite widespread interest in using human induced pluripotent stem cells (hiPSCs) in neurological disease modeling, a suitable model system to study human neuronal connectivity is lacking. Here, we report a comprehensive and efficient differentiation paradigm for hiPSCs that generate multiple CA3 pyramidal neuron subtypes as detected by single-cell RNA sequencing (RNA-seq). This differentiation paradigm exhibits characteristics of neuronal network maturation, and rabies virus tracing revealed synaptic connections between stem cell-derived dentate gyrus (DG) and CA3 neurons in vitro recapitulating the neuronal connectivity within the hippocampus. Because hippocampal dysfunction has been implicated in schizophrenia, we applied DG and CA3 differentiation paradigms to schizophrenia-patient-derived hiPSCs. We detected reduced activity in DG-CA3 co-culture and deficits in spontaneous and evoked activity in CA3 neurons from schizophrenia-patient-derived hiPSCs. Our approach offers critical insights into the network activity aspects of schizophrenia and may serve as a promising tool for modeling diseases with hippocampal vulnerability.

Keywords

hippocampus
CA3
DG
rabies tracing
pyramidal neurons
synaptic connectivity
single cell sequencing
neuronal diversity
disease-in-a-dish
schizophrenia

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Present address: Department of Hematology-Oncology, University of Freiburg, 79106 Freiburg, Germany

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