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

STAR Protoc. 2021 Apr 5;2(2):100425. doi: 10.1016/j.xpro.2021.100425. eCollection 2021 Jun 18.

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).

Keywords: Cell Biology; Flow Cytometry/Mass Cytometry; Neuroscience; Single Cell; Stem Cells.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Culture Techniques
  • Cell Line
  • Ependyma / cytology*
  • Female
  • Flow Cytometry / methods*
  • Gene Expression Profiling
  • Male
  • Mice
  • Mice, Inbred C57BL
  • Neural Stem Cells / cytology*
  • Single-Cell Analysis / methods*
  • Transcriptome / genetics*