Objective To obtain venous endothelial cells to be used in the research of blood vessel tissue engineering.Methods The external jugular vein of canine was resected under aseptic conditions.The primary endothelial cells were harvested,cultured,identified and multiplicated,and observed by optical microscopy.Result The endothelial cells of the canine external jugular vein,cultured and multiplicated in vitro,not only had good bioactivity,but also met the need of experiments in amount.Conclution Endothelial cells of canine external jugular vein cultured in vitro may fit the need of blood vessel tissue engineering.