In this study, to improve efficiency and functionality of cell-based biosensor microarrays, we introduced nanotopographical features on cell adhesive region. Micropatterned surface was created by poly (ethylene glycol) (PEG) hydrogel microstructures via photolithography. Nano-topographical features were introduced using polymeric nanofibers which were produced by electrospinning technique. Combining PEG hydrogel microstructure and polymeric nanofibers created a clear contrast between adhesion resist hydrogel walls and adhesion promoting nanofiber surfaces. When mammalian cells were seeded onto micropatterned nanostructure, cells only selectively adhered to nanofiber maintaining their viability, while adherent cells were not present on the hydrogel wall.