Isolation and culture of neural stem cells from adult mouse subventricular zone for genetic and pharmacological treatments with proliferation analysis

Daniel Z Radecki; Jayshree Samanta

doi:10.1016/j.xpro.2022.101153

Isolation and culture of neural stem cells from adult mouse subventricular zone for genetic and pharmacological treatments with proliferation analysis

STAR Protoc. 2022 Feb 3;3(1):101153. doi: 10.1016/j.xpro.2022.101153. eCollection 2022 Mar 18.

Authors

Daniel Z Radecki^{1

2}, Jayshree Samanta^{1

2}

Affiliations

¹ Department of Comparative Biosciences, School of Veterinary Medicine, University of Wisconsin-Madison, 2015 Linden Drive, Madison, WI 53706, USA.
² Stem Cell and Regenerative Medicine Center, University of Wisconsin-Madison, 2015 Linden Drive, Madison, WI 53706, USA.

Abstract

Neural stem cells (NSCs) from the subventricular zone (SVZ) of the mouse brain can be expanded in vitro and grown as neurospheres, which can be stored long-term in liquid nitrogen. Here, we present a protocol for isolation and culture of NSCs from the adult mouse SVZ. We describe how to grow and expand primary NSCs to neurospheres, followed by differentiation and nucleofection/pharmacological treatments. Finally, we describe RNA extraction, EdU labeling of the cells, and immunofluorescent analysis to examine their proliferation. For complete details on the use and execution of this protocol, please refer to Radecki et al. (2020).

Keywords: Cell Biology; Cell culture; Cell isolation; Neuroscience; Stem Cells.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Cell Differentiation / genetics
Cell Proliferation
Lateral Ventricles*
Mice
Neural Stem Cells*

Abstract

Publication types

MeSH terms

Grants and funding