Format

Send to

Choose Destination
Cell Rep. 2018 May 22;23(8):2509-2523. doi: 10.1016/j.celrep.2018.04.066.

Combining NGN2 Programming with Developmental Patterning Generates Human Excitatory Neurons with NMDAR-Mediated Synaptic Transmission.

Author information

1
Stanley Center for Psychiatric Research, Broad Institute of Harvard and MIT, Cambridge, MA 02142, USA; Department of Stem Cell and Regenerative Biology, Harvard University, Cambridge, MA 02138, USA.
2
Stanley Center for Psychiatric Research, Broad Institute of Harvard and MIT, Cambridge, MA 02142, USA.
3
Novartis Institutes for Biomedical Research, Novartis, Cambridge, MA 02139, USA.
4
Department of Stem Cell and Regenerative Biology, Harvard University, Cambridge, MA 02138, USA.
5
Umea University, Faculty of Medicine, Department of Clinical Sciences, Psychiatry, 901 85 Umea, Sweden.
6
Center for Human Genetic Research, Massachusetts General Hospital, Boston, MA 02114, USA.
7
Novartis Institutes for Biomedical Research, Novartis, 4056 Basel, Switzerland.
8
Stanley Center for Psychiatric Research, Broad Institute of Harvard and MIT, Cambridge, MA 02142, USA; McGovern Institute for Brain Research in the Department of Brain and Cognitive Sciences at MIT, Cambridge, MA 02139, USA.
9
Stanley Center for Psychiatric Research, Broad Institute of Harvard and MIT, Cambridge, MA 02142, USA; Department of Stem Cell and Regenerative Biology, Harvard University, Cambridge, MA 02138, USA. Electronic address: eggan@mcb.harvard.edu.

Abstract

Transcription factor programming of pluripotent stem cells (PSCs) has emerged as an approach to generate human neurons for disease modeling. However, programming schemes produce a variety of cell types, and those neurons that are made often retain an immature phenotype, which limits their utility in modeling neuronal processes, including synaptic transmission. We report that combining NGN2 programming with SMAD and WNT inhibition generates human patterned induced neurons (hpiNs). Single-cell analyses showed that hpiN cultures contained cells along a developmental continuum, ranging from poorly differentiated neuronal progenitors to well-differentiated, excitatory glutamatergic neurons. The most differentiated neurons could be identified using a CAMK2A::GFP reporter gene and exhibited greater functionality, including NMDAR-mediated synaptic transmission. We conclude that utilizing single-cell and reporter gene approaches for selecting successfully programmed cells for study will greatly enhance the utility of hpiNs and other programmed neuronal populations in the modeling of nervous system disorders.

KEYWORDS:

AMPAR; CAMK2A; NGN2; NMDAR; Wnt inhibition; dual SMAD inhibition; excitatory neurons; human stem cell; neuronal differentiation; single cell profiling

PMID:
29791859
PMCID:
PMC6003669
DOI:
10.1016/j.celrep.2018.04.066
[Indexed for MEDLINE]
Free PMC Article

Supplemental Content

Full text links

Icon for Elsevier Science Icon for PubMed Central
Loading ...
Support Center