An effective method of isolating endothelial cells from intact rat aorta

Rat aorta has been widely utilized in studies on cardiovascular diseases. Due to its small size, the endothelial cells are difficult to isolate. Here, we developed a modified method to obtain the endothelial cells from the rat aorta. The thoracic aorta of Wistar rat was harvested with its inside was turned out to expose the tunica intima. With its both ends ligated and cultured in ECM in T25 flask, a small amount of cells grew out from the aorta on day 4 and manifested the characteristic cobble stone-like monolayer on day 9-10. The cells were identified to be endothelial cells using reverse transcription (RT)-polymerase chain reaction (PCR), immunocytochemical staining for Von Willebrand Factor (vWF), Dil-Ac-LDL uptake assay, and capillary tube formation assay. This method serves as an easy way to get a large amount of endothelial cells from small-sized vessels, which, therefore, will greatly benefit the researches of vascular disease.