Objective To develop an innovative device for endothelial cell culture in vitro, namely, to develop a vascular endothelial cell culture device based on hemodynamic environment, so as to introduce the development and experimental study of endothelial cell culture device in vitro. Methods A device of dynamic culture system for endothelial cells in vitro on the basis of the existing research was designed with the theory and method of hemodynamics. The shear stress, positive stress and tensile stress existed at the same time in the flow environment. The development and experimental research of the device were described in detail from 5 aspects, such as the development background, structure and composition, design principle, theoretical basis and experimental research. Results The device could accurately simulate the hemodynamic environment of endothelial cells at normal level, with precise control of shear stress in 0-12 Pa range, positive stress in 0-15.96 kPa range, and tensile stress in 0-0.5 MPa range. Conclusions The device can provide a hemodynamic environment which is closer to the physiological conditions of human body, as well as a more ideal experimental environment and means for further exploring the mechanism of vascular intimal injury.