Efficient Generation of CA3 Neurons from Human Pluripotent Stem Cells Enables Modeling of Hippocampal Connectivity In Vitro

Anindita Sarkar, Arianna Mei, Apua C.M. Paquola, Shani Stern, Cedric Bardy, Jason R. Klug, Stacy Kim, Neda Neshat, Hyung Joon Kim, Manching Ku, Maxim N. Shokhirev, David H. Adamowicz, Maria C. Marchetto, Roberto Jappelli, Jennifer A. Erwin, Krishnan Padmanabhan, Matthew Shtrahman, Xin Jin, Fred H. Gage

Research output: Contribution to journalArticle

39 Scopus citations

Abstract

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. Video Abstract: [Figure presented] Sarkar et al. established a differentiation paradigm that generates human CA3 pyramidal neurons from ESCs and iPSCs and recapitulates hippocampal connectivity in vitro. This work reveals reduced levels of activity of schizophrenia-patient-derived neurons, offering opportunities for modeling diseases with hippocampal vulnerability.

Original languageEnglish (US)
Pages (from-to)684-697.e9
JournalCell Stem Cell
Volume22
Issue number5
DOIs
StatePublished - May 3 2018

Keywords

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

ASJC Scopus subject areas

  • Molecular Medicine
  • Genetics
  • Cell Biology

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    Sarkar, A., Mei, A., Paquola, A. C. M., Stern, S., Bardy, C., Klug, J. R., Kim, S., Neshat, N., Kim, H. J., Ku, M., Shokhirev, M. N., Adamowicz, D. H., Marchetto, M. C., Jappelli, R., Erwin, J. A., Padmanabhan, K., Shtrahman, M., Jin, X., & Gage, F. H. (2018). Efficient Generation of CA3 Neurons from Human Pluripotent Stem Cells Enables Modeling of Hippocampal Connectivity In Vitro. Cell Stem Cell, 22(5), 684-697.e9. https://doi.org/10.1016/j.stem.2018.04.009