Protocol for neurogenin-2-mediated induction of human stem cell-derived neural progenitor cells

Ellen J. Guss; Laila Sathe; Alexander Dai; Tim Derebenskiy; Ana Rodriguez Vega; Kevin Eggan; Michael F. Wells

STAR Protocols (Mar 2024)

Protocol for neurogenin-2-mediated induction of human stem cell-derived neural progenitor cells

Ellen J. Guss,
Laila Sathe,
Alexander Dai,
Tim Derebenskiy,
Ana Rodriguez Vega,
Kevin Eggan,
Michael F. Wells

Affiliations

Ellen J. Guss: Department of Stem Cell and Regenerative Biology, and Harvard Stem Cell Institute, Harvard University, Cambridge, MA 02138, USA
Laila Sathe: Department of Human Genetics, David Geffen School of Medicine at the University of California Los Angeles, Los Angeles, CA 90095, USA
Alexander Dai: Department of Human Genetics, David Geffen School of Medicine at the University of California Los Angeles, Los Angeles, CA 90095, USA
Tim Derebenskiy: Department of Human Genetics, David Geffen School of Medicine at the University of California Los Angeles, Los Angeles, CA 90095, USA
Ana Rodriguez Vega: Department of Human Genetics, David Geffen School of Medicine at the University of California Los Angeles, Los Angeles, CA 90095, USA
Kevin Eggan: Department of Stem Cell and Regenerative Biology, and Harvard Stem Cell Institute, Harvard University, Cambridge, MA 02138, USA; Stanley Center for Psychiatric Research, Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA
Michael F. Wells: Department of Human Genetics, David Geffen School of Medicine at the University of California Los Angeles, Los Angeles, CA 90095, USA; Molecular Biology Institute, University of California Los Angeles, Los Angeles, CA 90095, USA; Corresponding author

Journal volume & issue: Vol. 5, no. 1
p. 102878

Abstract

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Summary: Human pluripotent stem cell-derived neural progenitor cells (NPCs) are an essential tool for the study of brain development and developmental disorders such as autism. Here, we present a protocol to generate NPCs rapidly and reproducibly from human stem cells using dual-SMAD inhibition coupled with a brief pulse of mouse neurogenin-2 (Ngn2) overexpression. We detail the 48-h induction scheme deployed to produce these cells—termed stem cell-derived Ngn2-accelerated progenitor cells—followed by steps for expansion, purification, banking, and quality assessment.For complete details on the use and execution of this protocol, please refer to Wells et al.1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.

Published in STAR Protocols

ISSN: 2666-1667 (Online)
Publisher: Elsevier
Country of publisher: United States
LCC subjects: Science: Science (General)
Website: https://star-protocols.cell.com/

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Abstract

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