Isolation of mouse coronary endothelial cells

Shizhen Luo, Aaron H. Truong, Ayako Makino

Research output: Contribution to journalArticle

5 Scopus citations

Abstract

Endothelial cells line the inner wall of blood vessels and play an important role in the regulation of vascular tone, vascular permeability, and new vascular formation. Endothelial cell dysfunction is implicated in the development and progression of many cardiovascular diseases including ischemic heart disease. To examine the function and characterization of coronary endothelial cells, cell isolation is the first step and it requires high purity and quantity to conduct subsequent experiments. This protocol describes an efficient method to isolate adult mouse coronary endothelial cells. The mouse heart is dissected and minced into small pieces. After the digestion of the heart using dispase and collagenase II, cells are washed and incubated with magnetic beads which are conjugated with anti-CD31 antibody. The beads with endothelial cells are washed several times and are ready to use in various applications, including imaging and molecular biological experiments. Efficient isolation yields approximately 104 cells per one heart with over 90% purity.

Original languageEnglish (US)
Article numbere53985
JournalJournal of Visualized Experiments
Volume2016
Issue number113
DOIs
StatePublished - Jul 3 2016

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Keywords

  • CD31
  • Cellular biology
  • Dissection
  • Enzymatic digestion
  • Hearts
  • Issue 113
  • MCEC
  • Magnetic beads

ASJC Scopus subject areas

  • Neuroscience(all)
  • Chemical Engineering(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Immunology and Microbiology(all)

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