Synthetic substrates able to prevent the adhesion of cells, commonly referred to as nonfouling surfaces, are useful in a wide range of applications, including the culture of multicellular spheroids and the engineering of complex cell sheets. Nonfouling properties are commonly conferred to surfaces through superhydrophilic polymer-based coatings. At the same time, this type of treatment requires difficult, expensive, and time-consuming procedures, leading to high prices of the final product. In this perspective, the development of quick single-step methods to coat surfaces with nonfouling polymers would dramatically reduce the costs and increase the flexibility of such products. Moreover, this strategy would enable the process to be carried out directly by the end user. In this work, we report a fast and easy-to-use coating method to prevent the nonspecific adhesion of cells on tissue culture polystyrene (TCPS) surfaces. Our approach is based on the adsorption of poly(styrene-co-3-sulfopropyl methacrylate) copolymers comprising nonfouling hydrophilic moieties as well as functionalities that enhance the polymer adhesion to the substrate. The polymer adsorption is obtained from an aqueous mixture with a procedure that can be conveniently performed in short time. The kinetic of polymer adsorption as well as the effect of the polymer concentration was studied in order to reduce the time and cost of the entire procedure. Additionally, the polymer composition and the polymer density were optimized to completely avoid the adhesion of three different adherent cell lines, that is, CHO, A375-P, and HFF-1 cells.