title: Isolation and Culture of Aortic Smooth Muscle Cells and In Vitro Calcification Assay.
abstract: Elevated serum phosphorus is a major risk factor for vascular calcification, which is characterized by the presence of calcium phosphate deposits, mainly hydroxyapatite crystals. In vitro studies of phosphate-induced calcification show that vascular smooth muscle cells undergo calcification with features similar to those observed in pathological vascular calcification in vivo, including the presence of hydroxyapatite crystals. Here, we describe the double-collagenase digestion method for isolating vascular smooth muscle cells from aorta, and a method for inducing calcification in vitro using high phosphate concentration.
authors.affiliation: Laboratory of Molecular and Genetic Cardiovascular Pathophysiology, Vascular Biology Program, Centro Nacional de Investigaciones Cardiovasculares Carlos III (CNIC), Melchor Fernández Almagro 3, 28029, Madrid, Spain. metabol@hotmail.com.
authors.forename: Ricardo
authors.initials: R
authors.lastname: Villa-Bellosta
authors.affiliation: Laboratory of Molecular and Genetic Cardiovascular Pathophysiology, Vascular Biology Program, Centro Nacional de Investigaciones Cardiovasculares Carlos III (CNIC), Melchor Fernández Almagro 3, 28029, Madrid, Spain.
authors.forename: Magda R
authors.initials: MR
authors.lastname: Hamczyk
mesh_terms: D000818:Animals; D001011:Aorta / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D001018:Aortic Diseases / Q000139:chemically induced / Q000378:metabolism / Q000473:pathology*; D002130:Calcium Phosphates / Q000633:toxicity; D002469:Cell Separation / Q000379:methods*; D002478:Cells, Cultured; D017364:Collagenases / Q000378:metabolism; D003460:Crystallization; D017886:Durapatite / Q000378:metabolism; D051379:Mice; D009131:Muscle, Smooth, Vascular / Q000187:drug effects / Q000378:metabolism / Q000473:pathology*; D032389:Myocytes, Smooth Muscle / Q000187:drug effects / Q000378:metabolism / Q000473:pathology*; D061251:Primary Cell Culture; D061205:Vascular Calcification / Q000139:chemically induced / Q000378:metabolism / Q000473:pathology*
chemical_list: D002130:Calcium Phosphates; D017886:Durapatite; C020243:calcium phosphate; D017364:Collagenases
keywords: Aorta; Calcium phosphate deposits; Collagenase; Hydroxyapatite; Pi-induced calcification; Vascular smooth muscle cells