Villa-Bellosta Ricardo, Hamczyk Magda R
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.
Methods Mol Biol. 2015;1339:119-29. doi: 10.1007/978-1-4939-2929-0_8.
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.
血清磷升高是血管钙化的主要危险因素，其特征是存在磷酸钙沉积物，主要是羟基磷灰石晶体。磷酸盐诱导钙化的体外研究表明，血管平滑肌细胞会发生钙化，其特征与体内病理性血管钙化中观察到的特征相似，包括存在羟基磷灰石晶体。在此，我们描述了从主动脉中分离血管平滑肌细胞的双胶原酶消化法，以及使用高磷酸盐浓度在体外诱导钙化的方法。
