Non-thermal plasmas are emerging as a novel tool for the treatment of living tissues for biological and medical purpose. In this study, we described the effect of 4 min dielectric barrier discharge (DBD) plasma on both T98G cancer and HEK normal cell lines in the presence of different concentrations of osmolytes. This treatment strategy shows a specific inhibitory effect of a 240 s plasma exposure in the presence of osmolytes against T98G brain cancer cells only, but not on HEK normal cells. Based on these interesting properties of osmolytes, a non-thermal plasma appears to be a potential anticancer treatment strategy for different kinds of cancers in the presence of osmolytes.