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.