- 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
PMID33899012
RESUMEN
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).
MATERIALES
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Sigma-Aldrich
Disolución salina tamponada con fosfatos de Dulbecco, 10×, Modified, without calcium chloride and magnesium chloride, liquid, sterile-filtered, suitable for cell culture
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
Seroalbúmina bovina, heat shock fraction, protease free, suitable for hybridization, pH 7, ≥98%