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

Overview

Journal Cell Rep

Publisher Cell Press

Specialties Biology
Cell Biology
Molecular Biology

Date 2018 May 24

PMID 29791859

Citations 132

Authors

Ralda Nehme

Emanuela Zuccaro

Sulagna Dia Ghosh

Chenchen Li

John L Sherwood

Olli Pietilainen

Lindy E Barrett

Francesco Limone

Kathleen A Worringer

Sravya Kommineni

Ying Zang

Davide Cacchiarelli

Alex Meissner

Rolf Adolfsson

Stephen Haggarty

Jon Madison

Matthias Muller

Paola Arlotta

Zhanyan Fu

Guoping Feng

Kevin Eggan

Affiliations

Soon will be listed here.

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.

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