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Merck

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
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).

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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
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DAPI, for nucleic acid staining
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Trypsin inhibitor from Glycine max (soybean), powder, BioReagent, suitable for cell culture
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Ethylenediaminetetraacetic acid tetrasodium salt dihydrate, BioReagent, suitable for cell culture, 98.5-102.0%
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Seroalbúmina bovina, heat shock fraction, protease free, suitable for hybridization, pH 7, ≥98%