Protocols for endothelial cell isolation from mouse tissues: brain, choroid, lung, and muscle

STAR Protoc. 2021 Sep 14;2(3):100508. doi: 10.1016/j.xpro.2021.100508. eCollection 2021 Sep 17.

Abstract

Endothelial cells (ECs) harbor distinct phenotypical and functional characteristics depending on their tissue localization and contribute to brain, eye, lung, and muscle diseases such as dementia, macular degeneration, pulmonary hypertension, and sarcopenia. To study their function, isolation of pure ECs in high quantities is crucial. Here, we describe protocols for rapid and reproducible blood vessel EC purification established for scRNA sequencing from murine tissues using mechanical and enzymatic digestion followed by magnetic and fluorescence-activated cell sorting. For complete details on the use and execution of these protocol, please refer to Kalucka et al. (2020), Rohlenova et al. (2020), and Goveia et al. (2020).

Keywords: Cell isolation; Flow Cytometry/Mass Cytometry; Single Cell.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Brain / cytology*
  • Choroid / cytology*
  • Endothelial Cells / cytology*
  • Flow Cytometry / methods
  • Lung / cytology*
  • Male
  • Mice
  • Mice, Inbred C57BL
  • Muscles / cytology*