The utilization of non-biofouling poly(oligo(ethylene glycol) methacrylate) (pOEGMA) films as a background material for the generation of neuronal patterns is reported here. Our previously reported method, which was surface-initiated, atom transfer radical polymerization of OEGMA, and subsequent activation of terminal hydroxyl groups of pOEGMA with disuccinimidyl carbonate, was employed for the generation of activated pOEGMA films on glass. Poly-L-lysine was then microcontact-printed onto the activated polymer films, followed by backfilling with poly(ethylene glycol) moieties. E18 hippocampal neurons were cultured on the chemically patterned substrate, and the resulting neuronal networks were analyzed by phase-contrast microscopy and whole-cell patch clamp method. The results indicated that the pOEGMA films played an important role in the generation of good-quality neuronal patterns for up to two weeks without any negative effects to neurons.