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  • Cell population analysis of the adult murine subependymal neurogenic lineage by flow cytometry.

Cell population analysis of the adult murine subependymal neurogenic lineage by flow cytometry.

STAR protocols (2021-04-27)
Germán Belenguer, Pere Duart-Abadia, Ana Domingo-Muelas, Jose Manuel Morante-Redolat, Isabel Fariñas
ABSTRACT

This protocol provides a flow-cytometry-based procedure to classify and isolate all cells of the adult rodent subependymal zone (SEZ) neurogenic lineage, without the need for reporter mice, into different cell populations, including three neural stem cell (NSC) fractions with molecular signatures that are coherent with single-cell transcriptomics. Additionally, their cycling behavior can be assessed by means of 5-ethynyl-2'-deoxyuridine (EdU) incorporation. Our method allows the isolation of different NSC fractions and the functional assay of their cycling heterogeneity and quiescence-activation transitions. For complete details on the use, execution, and outcomes of this protocol, please refer to Belenguer et al. (2021).

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Trypsin inhibitor from Glycine max (soybean), powder, BioReagent, suitable for cell culture
Sigma-Aldrich
Ethylenediaminetetraacetic acid tetrasodium salt dihydrate, BioReagent, suitable for cell culture, 98.5-102.0%
Sigma-Aldrich
Bovine Serum Albumin, heat shock fraction, protease free, suitable for hybridization, pH 7, ≥98%
Sigma-Aldrich
Dulbecco′s Phosphate Buffered Saline, 10×, Modified, without calcium chloride and magnesium chloride, liquid, sterile-filtered, suitable for cell culture
Sigma-Aldrich
DAPI, for nucleic acid staining