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

doi:10.1016/j.xpro.2024.102878

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

STAR Protoc. 2024 Mar 15;5(1):102878. doi: 10.1016/j.xpro.2024.102878. Epub 2024 Feb 8.

Authors

Ellen J Guss¹, Laila Sathe², Alexander Dai², Tim Derebenskiy², Ana Rodriguez Vega², Kevin Eggan³, Michael F Wells⁴

Affiliations

¹ Department of Stem Cell and Regenerative Biology, and Harvard Stem Cell Institute, Harvard University, Cambridge, MA 02138, USA.
² Department of Human Genetics, David Geffen School of Medicine at the University of California Los Angeles, Los Angeles, CA 90095, USA.
³ 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.
⁴ 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. Electronic address: mfwells@mednet.ucla.edu.

Abstract

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.

Keywords: Cell culture; Neuroscience; Stem Cells.

MeSH terms

Animals
Cell Differentiation / physiology
Humans
Mice
Neural Stem Cells*
Pluripotent Stem Cells*

Grants and funding

T32 HG002536/HG/NHGRI NIH HHS/United States