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Merck

Protocol for mouse adult neural stem cell isolation and culture.

STAR protocols (2021-05-25)
Ahmed K M A Ahmed, Toke Jost Isaksen, Toshihide Yamashita
ABSTRACT

This protocol entails a simple method for isolation, culturing, and in vitro differentiation of adult neural stem cells from the dentate gyrus in the hippocampus and the subventricular zone of adult mice. Cultured adult neural stem cells are an important in vitro model to investigate stem cell properties such as proliferation and differentiation and to expand the understanding of plasticity in the adult brain. For complete details on the use and execution of this protocol, please refer to Isaksen et al. (2020).

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Monoclonal Anti-MAP2 antibody produced in mouse, ascites fluid, clone HM-2
Sigma-Aldrich
Hydrocortisone, BioReagent, suitable for cell culture
Sigma-Aldrich
Sodium selenite, BioReagent, suitable for cell culture, ≥98%
Sigma-Aldrich
HEPES, ≥99.5% (titration)
Sigma-Aldrich
Trypsin inhibitor from Glycine max (soybean), powder, BioReagent, suitable for cell culture
Sigma-Aldrich
Anti-SOX-2 Antibody, clone 6F1.2, Chemicon®, from mouse
Sigma-Aldrich
Anti-Nestin Antibody, clone rat-401, clone rat-401, Chemicon®, from mouse
Sigma-Aldrich
Biotin, ≥99% (HPLC), lyophilized powder
Sigma-Aldrich
Forskolin, For use in molecular biology applications
Sigma-Aldrich
Anti-Olig2 Antibody, clone 211F1.1, clone 211F1.1, from mouse