Cardiac microvascular endothelial cells express α-smooth muscle actin and show low NOS III activity
- 1 May 1999
- journal article
- research article
- Published by American Physiological Society in American Journal of Physiology-Heart and Circulatory Physiology
- Vol. 276 (5), H1755-H1768
- https://doi.org/10.1152/ajpheart.1999.276.5.h1755
Abstract
We established a culture system of porcine coronary microvascular endothelial cells (MVEC) with high cellular yield and purity >98%. Endothelial origin was confirmed by immunostaining, immunoblotting and fluorescence-activated cell sorter (FACS) analysis using low-density lipoprotein uptake, CD31, von Willebrand factor, and the lectin Dolichos biflorus agglutinin. MVEC were positive for α-smooth muscle actin in culture and in myocardium, as confirmed by FACS. Of the primary MVEC, ∼30% expressed nitric oxide synthase (NOS) III in numbers decreasing from the first passage (6 ± 1%) to the second passage (4 ± 1%; P < 0.001 vs. primary isolates), whereas ∼100% of aortic endothelial cells (AEC) expressed NOS III. In AEC, NOS III activity (pmol citrulline ⋅ mg protein−1⋅ min−1) was 80 ± 10 and was nearly abolished in the absence of calcium (5 ± 1, P < 0.001). In primary MVEC, however, NOS III activity in the presence and absence of calcium was 20 ± 4 and 25 ± 5, respectively. We conclude that cardiac MVEC, in contrast to AEC, contain α-smooth muscle actin, show low-grade NOS III activity, and provide a suitable in vitro system for the study of endothelial pathophysiology.Keywords
This publication has 33 references indexed in Scilit:
- Heterogeneity of Endothelial CellsArteriosclerosis, Thrombosis, and Vascular Biology, 1997
- Cardiac endothelium and tissue growthProgress in Cardiovascular Diseases, 1997
- Endothelial Nitric Oxide Synthase Targeting to CaveolaeJournal of Biological Chemistry, 1996
- Molecular Regulation of the Bovine Endothelial Cell Nitric Oxide Synthase by Transforming Growth Factor–β 1Arteriosclerosis, Thrombosis, and Vascular Biology, 1995
- Regulation of Endothelial Constitutive Nitric Oxide Synthase by Protein Kinase CHypertension, 1995
- Growth and Physiological Functions of Vascular Endothelial Cells in a New Serum-Free Medium (SFM)Experimental Cell Research, 1993
- In situ localization of transforming growth factor β1 in porcine heart: Enhanced expression after chronic coronary artery constrictionJournal of Molecular and Cellular Cardiology, 1991
- Response of the coronary vasculature to myocardial hypertrophyJournal of the American College of Cardiology, 1990
- Differential distribution of von Willebrand factor in endothelial cells. Comparison between normal pigs and pigs with von Willebrand disease.Arteriosclerosis: An Official Journal of the American Heart Association, Inc., 1987
- Identification and isolation of endothelial cells based on their increased uptake of acetylated-low density lipoprotein.The Journal of cell biology, 1984