Preparation of single cell suspensions enriched in mouse brain vascular cells for single-cell RNA sequencing

STAR Protoc. 2021 Aug 5;2(3):100715. doi: 10.1016/j.xpro.2021.100715. eCollection 2021 Sep 17.

Abstract

Cerebral blood vessels supply oxygen and nutrients, remove metabolic waste, and play a critical role in maintaining brain homeostasis. Cerebrovasculature is composed of heterogeneous populations of brain vascular cells (BVCs). A major challenge in effective cerebrovascular transcriptional profiling is high-quality BVC procurement, permitting high sequencing depth. Here, we establish cell isolation procedures for glio-vascular cell-enriched single-cell RNA sequencing enabling unbiased characterization of BVC transcriptional heterogeneity. Our approach can be used to address vascular-specific contribution to brain diseases. For complete details on the use and execution of this protocol, please refer to Yamazaki et al. (2021).

Keywords: Cell isolation; Neuroscience; RNAseq; Single Cell.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Blood Cells / metabolism
  • Blood Vessels
  • Brain / blood supply*
  • Brain / metabolism
  • Cardiovascular System / metabolism
  • Cerebrovascular Circulation / physiology
  • Exome Sequencing / methods*
  • Mice
  • Sequence Analysis, RNA / methods
  • Single-Cell Analysis / methods*
  • Suspensions / isolation & purification

Substances

  • Suspensions