The short-term interactions of chondrosarcoma cells with polyelectrolyte multilayer films built up by the alternate adsorption of poly(L-lysine) (PLL) and poly(L-glutamic acid) (PGA) was studied in the presence and in the absence of serum. The films and their interaction with serum proteins were first characterized by means of optical waveguide lightmode spectroscopy, quartz crystal microbalance, and zeta potential measurements. In a serum-containing medium, the detachment forces measured by the micropipet technique were about eight times smaller on PGA-ending than on PLL-ending films. For these latter ones, the adhesion force decreased when the film thickness increased. In a serum-free medium, the differences between the negative- and positive-ending films were enhanced: adhesion forces on PLL-ending films were 40-100% higher, whereas no cellular adherence was found on PGA-terminating films. PGA-ending films were found to prevent the adsorption of serum proteins, whereas important protein adsorption was always observed on PLL-ending films. These results show how cell interactions with polyelectrolyte films can be tuned by the type of the outermost layer, the presence of proteins, and the number of layers in the film.