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

Cell Rep
Authors
Keywords
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.

Year of Publication
2018
Journal
Cell Rep
Volume
23
Issue
8
Pages
2509-2523
Date Published
2018 05 22
ISSN
2211-1247
DOI
10.1016/j.celrep.2018.04.066
PubMed ID
29791859
PubMed Central ID
PMC6003669
Links
Grant list
U01 MH105669 / MH / NIMH NIH HHS / United States
U01 MH115727 / MH / NIMH NIH HHS / United States