Protocol for neurogenin-2-mediated induction of human stem cell-derived neural progenitor cells
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 coup...
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| Format: | Article |
| Language: | English |
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Elsevier
2024-03-01
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| Series: | STAR Protocols |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2666166724000431 |
| _version_ | 1797319435314266112 |
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| author | Ellen J. Guss Laila Sathe Alexander Dai Tim Derebenskiy Ana Rodriguez Vega Kevin Eggan Michael F. Wells |
| author_facet | Ellen J. Guss Laila Sathe Alexander Dai Tim Derebenskiy Ana Rodriguez Vega Kevin Eggan Michael F. Wells |
| author_sort | Ellen J. Guss |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-03-08T04:06:50Z |
| format | Article |
| id | doaj.art-b3cd8ded106c4302b87c41c20dab8227 |
| institution | Directory Open Access Journal |
| issn | 2666-1667 |
| language | English |
| last_indexed | 2024-03-08T04:06:50Z |
| publishDate | 2024-03-01 |
| publisher | Elsevier |
| record_format | Article |
| series | STAR Protocols |
| spelling | doaj.art-b3cd8ded106c4302b87c41c20dab82272024-02-09T04:49:15ZengElsevierSTAR Protocols2666-16672024-03-0151102878Protocol for neurogenin-2-mediated induction of human stem cell-derived neural progenitor cellsEllen J. Guss0Laila Sathe1Alexander Dai2Tim Derebenskiy3Ana Rodriguez Vega4Kevin Eggan5Michael F. Wells6Department of Stem Cell and Regenerative Biology, and Harvard Stem Cell Institute, Harvard University, Cambridge, MA 02138, USADepartment of Human Genetics, David Geffen School of Medicine at the University of California Los Angeles, Los Angeles, CA 90095, USADepartment of Human Genetics, David Geffen School of Medicine at the University of California Los Angeles, Los Angeles, CA 90095, USADepartment of Human Genetics, David Geffen School of Medicine at the University of California Los Angeles, Los Angeles, CA 90095, USADepartment of Human Genetics, David Geffen School of Medicine at the University of California Los Angeles, Los Angeles, CA 90095, USADepartment 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, USADepartment 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 authorSummary: 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.http://www.sciencedirect.com/science/article/pii/S2666166724000431Cell cultureNeuroscienceStem Cells |
| spellingShingle | Ellen J. Guss Laila Sathe Alexander Dai Tim Derebenskiy Ana Rodriguez Vega Kevin Eggan Michael F. Wells Protocol for neurogenin-2-mediated induction of human stem cell-derived neural progenitor cells STAR Protocols Cell culture Neuroscience Stem Cells |
| title | Protocol for neurogenin-2-mediated induction of human stem cell-derived neural progenitor cells |
| title_full | Protocol for neurogenin-2-mediated induction of human stem cell-derived neural progenitor cells |
| title_fullStr | Protocol for neurogenin-2-mediated induction of human stem cell-derived neural progenitor cells |
| title_full_unstemmed | Protocol for neurogenin-2-mediated induction of human stem cell-derived neural progenitor cells |
| title_short | Protocol for neurogenin-2-mediated induction of human stem cell-derived neural progenitor cells |
| title_sort | protocol for neurogenin 2 mediated induction of human stem cell derived neural progenitor cells |
| topic | Cell culture Neuroscience Stem Cells |
| url | http://www.sciencedirect.com/science/article/pii/S2666166724000431 |
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