title: Isolation and in vitro culture of vascular endothelial cells from mice.
abstract: In cardiovascular disorders, understanding of endothelial cell (EC) function is essential to elucidate the disease mechanism. Although the mouse model has many advantages for in vivo and in vitro research, efficient procedures for the isolation and propagation of primary mouse EC have been problematic. We describe a high yield process for isolation and in vitro culture of primary EC from mouse arteries (aorta, braches of superior mesenteric artery, and cerebral arteries from the circle of Willis). Mouse arteries were carefully dissected without damage under a light microscope, and small pieces of the vessels were transferred on/in a Matrigel matrix enriched with endothelial growth supplement. Primary cells that proliferated in Matrigel were propagated in advanced DMEM with fetal calf serum or platelet-derived serum, EC growth supplement, and heparin. To improve the purity of the cell culture, we applied shearing stress and anti-fibroblast antibody. EC were characterized by a monolayer cobble stone appearance, positive staining with acetylated low density lipoprotein labeled with 1,1'-dioctadecyl-3,3,3',3'-tetramethyl-indocarbocyanine perchlorate, RT-PCR using primers for von-Willebrand factor, and determination of the protein level endothelial nitric oxide synthase. Our simple, efficient method would facilitate in vitro functional investigations of EC from mouse vessels.
authors.affiliation: Department of Physiology, School of Medicine, Ewha Womans University, Seoul 157-710, Korea.
authors.forename: Shinkyu
authors.initials: S
authors.lastname: Choi
authors.affiliation: Department of Physiology, School of Medicine, Ewha Womans University, Seoul 157-710, Korea.
authors.forename: Ji Aee
authors.initials: JA
authors.lastname: Kim
authors.affiliation: Department of Thoracic & Cardiovascular Surgery and Ewha Womans University Global Top 5 Research Program, School of Medicine, Ewha Womans University, Seoul 157-710, Korea.
authors.forename: Kwan Chang
authors.initials: KC
authors.lastname: Kim
authors.affiliation: Department of Physiology, School of Medicine, Ewha Womans University, Seoul 157-710, Korea.
authors.forename: Suk Hyo
authors.initials: SH
authors.lastname: Suh
keywords: Endothelial cells; In vitro culture; Mouse vessels