title: Isolation of Mouse Coronary Endothelial Cells.
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 10(4) cells per one heart with over 90% purity.
authors.affiliation: Department of Physiology, University of Arizona, Tucson.
authors.forename: Shizhen
authors.initials: S
authors.lastname: Luo
authors.affiliation: Department of Physiology, University of Arizona, Tucson.
authors.forename: Aaron H
authors.initials: AH
authors.lastname: Truong
authors.affiliation: Department of Physiology, University of Arizona, Tucson; aymakino@email.arizona.edu.
authors.forename: Ayako
authors.initials: A
authors.lastname: Makino
mesh_terms: D000818:Animals; D002469:Cell Separation; D002478:Cells, Cultured; D042783:Endothelial Cells; D004730:Endothelium, Vascular; D006321:Heart; D018189:Immunomagnetic Separation; D051379:Mice