Analyzing mouse neural stem cell and progenitor cell proliferation using EdU incorporation and multicolor flow cytometry

Abstract

Summary This protocol describes an ex vivo approach to identify and quantify the proportions of proliferating neural stem cells and progenitors of the mouse subventricular zone. It uses ethynyl deoxyuridine (EdU) incorporation to identify dividing cells, combined with multicolor flow cytometry for 4 cell surface antigens to distinguish between 8 phenotypically distinct mouse neural progenitors and stem cells. It has been optimized for wild-type neonatal mice but can be used on mice of any postnatal age. For complete details on the use and execution of this profile, please refer to Kumari et al. (2020) .

Subventricular zone adult mouse neural stem cells require insulin receptor for self-renewal

“…The treated cells were propagated for an additional 5 days and then dissociated for flow cytometric analysis as outlined in Figure 1 A. Vehicle-treated cells were tdT negative, whereas tamoxifen-treated cells were strongly tdT positive ( Figures 1 B and B′). Using a flow cytometry panel established previously in our lab ( Buono et al., 2012 ; Velloso et al., 2022 ), we parsed cells within the neurosphere into NSCs and seven different intermediate progenitors ( Figure 1 C). This analysis revealed that the ΔNSC-IRKO spheres had 3.5-fold fewer NSCs compared with the ΔNSC-IR WT when only the tdT + cells were evaluated.…”

In the pursuit of new social neurons. Neurogenesis and social behavior in mice: A systematic review

“…These broad-spectrum variables are presented in articles that related social exploration and neurogenesis triggered by genetic mutations that affected numerous systems (Lehmann et al, 2013;Guha et al, 2014;Garrett et al, 2018;Alen et al, 2019;Zhang et al, 2019;Amar et al, 2021;Velloso et al, 2022), types of cells and proteins (Jenniches et al, 2016;Zhang et al, 2016;Salminen et al, 2017;Alén et al, 2019;Zhang et al, 2020;Nacer et al, 2021) plus…”