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 journalArticlepeer-review

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

Bibliographical note

Funding Information:
We thank Illumina (San Diego, CA) and Bio-Rad (CA) for early access of SureCell WTA 3? Library Prep Kit and the ddSEQ System. We thank Dr. Marilyn Farquhar for the use of the electron microscopy facility at the University of California, San Diego; Ying Jones, Jeffrey Chang, and Joan Valls for technical assistance; and Servier Medical Art and Jamie Simon for illustrations. We thank Mary Lynn Gage for editorial comments and Diana Yu and Ben Lacar for valuable comments on the project. This work was supported by CIRM (grant TG2-01158) (A.S.), the Streim Foundation (A.S.), the Helmsley Foundation (F.H.G.), the JPB Foundation (F.H.G.), the Engman Foundation (F.H.G.), the NIMH (grants U01-MH106882 and U19-MH106434) (F.H.G.), the NIMH (grants MH101634 and MH113924) (K.P.), a NARSAD Young Investigator Award (K.P.), the Razavi Newman Integrative Genomics and Bioinformatics Core Facility of the Salk Institute with funding from the NIH-NCI CCSG (grant P30 014195), and the Waitt Advanced Biophotonics Core Facility of the Salk Institute, with funding from the NIH-NCI CCSG (grant P30 014195) and NINDS (neuroscience core grant NS072031) and the Waitt Foundation.

Funding Information:
We thank Illumina (San Diego, CA) and Bio-Rad (CA) for early access of SureCell WTA 3′ Library Prep Kit and the ddSEQ System. We thank Dr. Marilyn Farquhar for the use of the electron microscopy facility at the University of California, San Diego; Ying Jones, Jeffrey Chang, and Joan Valls for technical assistance; and Servier Medical Art and Jamie Simon for illustrations. We thank Mary Lynn Gage for editorial comments and Diana Yu and Ben Lacar for valuable comments on the project. This work was supported by CIRM (grant TG2-01158) (A.S.), the Streim Foundation (A.S.), the Helmsley Foundation (F.H.G.), the JPB Foundation (F.H.G.), the Engman Foundation (F.H.G.), the NIMH (grants U01-MH106882 and U19-MH106434) (F.H.G.), the NIMH (grants MH101634 and MH113924) (K.P.), a NARSAD Young Investigator Award (K.P.), the Razavi Newman Integrative Genomics and Bioinformatics Core Facility of the Salk Institute with funding from the NIH-NCI CCSG (grant P30 014195), and the Waitt Advanced Biophotonics Core Facility of the Salk Institute, with funding from the NIH-NCI CCSG (grant P30 014195) and NINDS (neuroscience core grant NS072031) and the Waitt Foundation.

Funding Information:
We thank Illumina (San Diego, CA) and Bio-Rad (CA) for early access of SureCell WTA 3′ Library Prep Kit and the ddSEQ System. We thank Dr. Marilyn Farquhar for the use of the electron microscopy facility at the University of California, San Diego; Ying Jones, Jeffrey Chang, and Joan Valls for technical assistance; and Servier Medical Art and Jamie Simon for illustrations. We thank Mary Lynn Gage for editorial comments and Diana Yu and Ben Lacar for valuable comments on the project. This work was supported by CIRM (grant TG2-01158 ) (A.S.), the Streim Foundation (A.S.), the Helmsley Foundation (F.H.G.), the JPB Foundation (F.H.G.), the Engman Foundation (F.H.G.), the NIMH (grants U01-MH106882 and U19-MH106434 ) (F.H.G.), the NIMH (grants MH101634 and MH113924 ) (K.P.), a NARSAD Young Investigator Award (K.P.), the Razavi Newman Integrative Genomics and Bioinformatics Core Facility of the Salk Institute with funding from the NIH-NCI CCSG (grant P30 014195 ), and the Waitt Advanced Biophotonics Core Facility of the Salk Institute , with funding from the NIH-NCI CCSG (grant P30 014195 ) and NINDS (neuroscience core grant NS072031 ) and the Waitt Foundation .

Publisher Copyright:
© 2018 Elsevier Inc.

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

Fingerprint

Dive into the research topics of 'Efficient Generation of CA3 Neurons from Human Pluripotent Stem Cells Enables Modeling of Hippocampal Connectivity In Vitro'. Together they form a unique fingerprint.

Cite this