Transcription Factor–Mediated Differentiation of Human iPSCs into Neurons

Michael S. Fernandopulle; Ryan Prestil; Christopher Grunseich; Chao Wang; Li Gan; Michael E. Ward

doi:10.1002/cpcb.51

journal article May 18, 2018

Transcription Factor–Mediated Differentiation of Human iPSCs into Neurons

Michael S. Fernandopulle Ryan Prestil Christopher Grunseich Chao Wang

Li Gan

Michael E. Ward

Current Protocols in Cell Biology Vol. 79 No. 1 · Wiley

View at Publisher Save 10.1002/cpcb.51

Abstract

Abstract

Accurate modeling of human neuronal cell biology has been a long‐standing challenge. However, methods to differentiate human induced pluripotent stem cells (iPSCs) to neurons have recently provided experimentally tractable cell models. Numerous methods that use small molecules to direct iPSCs into neuronal lineages have arisen in recent years. Unfortunately, these methods entail numerous challenges, including poor efficiency, variable cell type heterogeneity, and lengthy, expensive differentiation procedures. We recently developed a new method to generate stable transgenic lines of human iPSCs with doxycycline‐inducible transcription factors at safe‐harbor loci. Using a simple two‐step protocol, these lines can be inducibly differentiated into either cortical (i
3
Neurons) or lower motor neurons (i
3
LMN) in a rapid, efficient, and scalable manner (Wang et al., 2017). In this manuscript, we describe a set of protocols to assist investigators in the culture and genetic engineering of iPSC lines to enable transcription factor–mediated differentiation of iPSCs into i
3
Neurons or i
3
LMNs, and we present neuronal culture conditions for various experimental applications. © 2018 by John Wiley & Sons, Inc.

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Details

Published: May 18, 2018
Vol/Issue: 79(1)
License: View

Authors

M

Michael S. Fernandopulle

National Institute of Neurological Disorders and Stroke, National Institutes of Health Bethesda Maryland

R

Ryan Prestil

National Institute of Neurological Disorders and Stroke, National Institutes of Health Bethesda Maryland

C

Christopher Grunseich

National Institute of Neurological Disorders and Stroke, National Institutes of Health Bethesda Maryland

C

Chao Wang

Gladstone Institute of Neurological Disease, Gladstone Institutes San Francisco California

L

Li Gan

Gladstone Institute of Neurological Disease, Gladstone Institutes San Francisco California

M

Michael E. Ward

National Institute of Neurological Disorders and Stroke, National Institutes of Health Bethesda Maryland

Cite This Article

Michael S. Fernandopulle, Ryan Prestil, Christopher Grunseich, et al. (2018). Transcription Factor–Mediated Differentiation of Human iPSCs into Neurons. Current Protocols in Cell Biology, 79(1). https://doi.org/10.1002/cpcb.51

Transcription Factor–Mediated Differentiation of Human iPSCs into Neurons

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