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通过流式细胞术分析成年鼠室管膜下神经发生谱系的细胞群体。

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

机构信息

Centro de Investigación Biomédica en Red sobre Enfermedades Neurodegenerativas (CIBERNED), Universidad de Valencia, 46100 Burjassot, Spain.

Instituto Universitario de Biotecnología y Biomedicina (BIOTECMED), Universidad de Valencia, 46100 Burjassot, Spain.

出版信息

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

DOI:10.1016/j.xpro.2021.100425
PMID:33899012
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC8056273/
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).

摘要

本方案提供了一种基于流式细胞术的方法，可将成年啮齿动物室管膜下区（SEZ）神经发生谱系的所有细胞无需报告基因小鼠，分类并分离为不同的细胞群，包括具有与单细胞转录组学一致的分子特征的三个神经干细胞（NSC）亚群。此外，还可以通过 5-乙炔基-2'-脱氧尿苷（EdU）掺入来评估其细胞周期行为。我们的方法允许分离不同的 NSC 亚群，并对其细胞周期异质性和静止-激活转变进行功能分析。如需详细了解该方案的使用、执行和结果，请参考 Belenguer 等人（2021 年）的研究。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/41f3/8056273/483b355c3fed/gr3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/41f3/8056273/824ebbec2e96/fx1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/41f3/8056273/53565e7104ea/gr1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/41f3/8056273/843fe04a17aa/gr2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/41f3/8056273/483b355c3fed/gr3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/41f3/8056273/824ebbec2e96/fx1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/41f3/8056273/53565e7104ea/gr1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/41f3/8056273/843fe04a17aa/gr2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/41f3/8056273/483b355c3fed/gr3.jpg

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