Abstract: Abstract: Objective To establish a simple and efficient method for isolating and culturing vascular endothelial cells of rat thoracic aorta. Methods Under aseptic condition, the thoracic aorta was harvested from healthy male Wistar rat. After stripping off the adventitia, the thoracic aorta were cut into vessel rings with length of 1.0～1.5mm. The vessel rings were placed on culture dish vertically and filled with the culture medium DMEM/ F12 containing 10 % fetal bovine serum; After 24h, the vessel rings had attached on the culture dish firmly and some new culture medium were added; 3～4d later, the thoracic aorta was discarded and the new culture medium was added into the culture dish. The migrating cells were digested by 0.25 % pancreatic enzyme for serial subcultivation. The endothelial cells were identified by morphological, immunohistochemical and flow-cytometry methods with anti-vWF antibody, which was regarded as the marker of endothelial cells. Results A amount of cells migrated from the aorta and adhered to the bottom of culture dish 3～4d after cultivation. The confluent cells grew rapidly after being digested with 0.25% pancreatic enzyme and showed the typical cobblestone appearance. The cells were identified as endothelial cells, its positive ratio was (98.3±0.2)% by using immunostaining and flow-cytometry with vWF. Conclusion The modified explanting cultivation method for isolating and culturing was established in the this study. This method is simple, easy to handle and more economical and applicable as does not need collagen and endothelial cell growth promoting substrate. In addition, this modified method is especially suitable for isolation and cultivation of vascular endothelial cells from small arteries.

Key words: Key words: endothelial cells, thoracic aorta, rats