Three-dimensional advance dynamic culture system promotes microvessel development from cultured endothelial cells in vitro

The present study describes a dynamic system that can be used in a three- dimensional (3D) in vitro cell culture environment which promotes new microvessel formation. Human umbilical vein endothelial cells (HUVECs) were used in this study to form the inner lining of the microvessel, guided by the dynamic flow produced by our system within a 3D matrix made from fibrin. A 2D environment was used as a comparison. The dynamic flow was set to produce 5, 10, 15 or 20 dynes/cm2 shear stress to the cells in culture. Cultured HUVECs were observed for the increase in cell numbers and formation of microvessels. Cells cultured after 2 days demonstrated an increase in cell numbers when subjected to 10dynes/cm2 or more. By day 4, cells appeared to have altered morphologies and were oriented towards the direction of fluid flow. From the fluorescence images observed, it became apparent that there were microvascular channels forming in the 3D cultures. Our dynamic flow system appears to influence endothelial cells to promote microvascular formations in a 3D environment