Risperidone is a commonly used antipsychotic drug in clinic. Previous studies have found that risperidone has a potential of enhancing differentiation of neural stem cells (NSCs). Malignant gliomas are associated with poor prognosis, largely due to the presence of glioma stem-like cells (GSLCs), which share many properties with adult NSCs. Hence, we aimed to investigate the effects of risperidone on the self-renewal and differentiation of GSLCs and the underlying mechanisms. Our data showed that risperidone inhibited self-renewal and induced differentiation of GSLCs into oligodendrocyte-like cells, which expressed typical markers for oligodendrocytes in vitro. Moreover, risperidone inhibited the GSLCs-initiated gliomagenesis in vivo. Risperidone treatment decreased activity of β-catenin, and increased intracellular Ca(2+) concentration, stromal interaction molecule 1 (STIM1), and calcium-sensing receptor (CaSR), suggesting the involvement of Wnt signaling. Taken together, our study suggests that risperidone may promote differentiation of glioma stem-like cells through the Wnt signaling pathway.