Nature uses vascular systems to permit large‐area control over the functionality of surfaces that lie above them. In this work, the application of this concept to the control of a hybrid living–nonliving system is demonstrated. Defined arrangements of vascular channels are created in agar using a fugitive ink printing method. The antibiotic gentamicin is then introduced into the vascular network where it diffuses to the surface and interacts with a model system of Escherichia coli cells. The cells either live or die depending on their distance from the underlying channels, permitting spatial control over the biological system. Using single‐channel systems to define critical parameters, a theoretical model is developed to define the final surface pattern based solely on the arrangement of the underlying vascular channels. The model is then successfully used to create more complex arrangements of cells at the surface. Finally, by introducing different types of active compounds into separate vascular channels, a mixture of bacterial species is separated and localized at defined points. This work demonstrates the ability of bioinspired embedded vascular systems to predictably control a biological system at a surface, laying the groundwork for future spatially and temporally controlled biointerfaces in both industry and medicine.