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

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

  • STAR Protoc. 2024 Feb 8;5(1):102878. doi: 10.1016/j.xpro.2024.102878.
Ellen J Guss 1 Laila Sathe 2 Alexander Dai 2 Tim Derebenskiy 2 Ana Rodriguez Vega 2 Kevin Eggan 3 Michael F Wells 4
Affiliations

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: [email protected].
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.

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