A simple photolithographic process for directly patterning glycidyl methacrylate modified hyaluronic acid features onto UV curable perfluoropolyether-based surfaces is presented. Due to the versatility of the developed method, HA spotted areas with different geometrical features could be rapidly and inexpensively designed. In addition, the excellent antifouling and fouling-release properties of the substrates enabled direct HA baits photo-grafting onto PFPEs without further surface passivation or chemical modification to avoid not specific adsorption. The aim of the study was to locally switch the surface properties of the PFPEs from cells and protein repulsive to adherent. Particularly, we exploited HA well-known preferential interactions with CD44 transmembrane receptors to selectively immobilize cancer cells. Living cell arrays offer a higher-resolution visualization of HA-CD44 interactions and may provide a deep insight into understanding molecular mechanisms needed to develop selective therapies and diagnosis against tumor growth.