Isolation and Culture of Endothelial Cells from Large Vessels

Abstract

The endothelium, which is at the interface between circulating blood and the vascular wall, comprises a simple squamous layer of cells that lines the inner surface of all blood vessels. Endothelial cells are highly metabolically active and play an important role in many physiological functions, including control of vasomotor tone, blood cell trafficking, vascular permeability, and maintenance of vascular integrity (Mensah, Vascul Pharmacol 46(5):310–314, 2007; Yetik-Anacak and Catravas, Vascul Pharmacol 45(5):268–276, 2006). Endothelial cells are characteristically ‘quiescent’ in that they do not actively proliferate, with the average lifespan of an endothelial cell being >1 year. The endothelium is very sensitive to mechanical stimuli (stretch, shear stress, pressure), humoral agents (angiotensin II (Ang II), endothelin-1 (ET-1), aldosterone, bradykinin, thromoxane) and chemical factors (glucose, reactive oxygen species (ROS)) and responds by releasing endothelial-derived mediators, such as nitric oxide (NO), prostacyclin (PGI2), platelet-activating factor (PAF), C-type atrial natriuretic peptide (ANP), and ET-1 to regulate vascular tone, prevent thrombosis and inflammation, and maintain structural integrity. Primary culture of endothelial cells is an important tool in dissecting the role of the endothelium in many physiological or pathological responses. This chapter describes the explant method for culture of endothelial cells from large vessels. Cells derived by the protocol described here can be used for cell biology and molecular biology studies in hypertension and other cardiovascular diseases where endothelial function may be impaired.