Shape-dependent endothelial cell properties that are independent of fluid shear stress remain largely undefined. More recently, surface micropatterning has been investigated as an approach to control the morphology and orientation of endothelial cells using synthetic substratum. Most research studies have reported the effects of microtopography on sub-confluent layers of cells, which has been the standard in research investigations to date. In this book, surface micropatterning is used to mimic the natural endothelial cell vasculature in confluent layers. Substrates with patterned microgrooves provide a method for evaluating the interplay between cell orientation and spatial localization of membrane proteins, which may be patterned using microcontact printing techniques.