[Culture and characterization of porcine coronary endothelial cells].

Abstract

Introduction: Vascular endothelium possesses biological properties that are involved in important physiological functions such vascular permeability, vascular tone regulation and angiogenesis. The difficulty of culture and long-term maintenance of sufficient amount of normal endothelial cells has proven to be the limitation for the understanding of the biological function of the endothelium. Therefore, the aim of this study was to culture and characterize the porcine coronary endothelial cells.

Material and methods: The endothelial cells were isolated by collagenase treatment and cultured in culture dishes coated with collagen, prepared from rat tail, containing medium RPMI1640/M199 and 15% fetal calf serum supplemented with antibiotics and fungizon. The cells were maintained to grown at 37 degrees C. The medium was changed one day after and then every two day. The cells were incubated with Dil-labeled-acetylated-LDL for determination of acetylated-LDL uptake. Confluence cultures of cells were examined by phase-contrast and confocal microscopy.

Results: After a day of culture, the endothelial cells adhere to the collagen and began to grow. While multiplying themselves, they colonize little by little the body of the surface of culture to form to confluence a monolayer of flat cells relatively homogenous. To confluence, the proliferation of the endothelial cells is inhibited by the contact and the cells present a polygonal aspect. Our results show that all the cultivated cells were strongly positive for acetylated-LDL markers. The endothelial cells, cultivated until the second passage corresponding to the second culture of the primary cultures, continued to present a good fluorescence.

Conclusion: Porcine coronary endothelial cells represent a useful in vitro model to study biological and physiopathological properties of vascular endothelium.