Previously, we found safrole oxide could promote VEC apoptosis, however, it is not known whether it can induce NSC apoptosis. It is reported that neural stem cells (NSCs) are localized in a vascular niche. But the effects of apoptosis in vascular endothelial cells (VEC) on NSC growth and differentiation are not clear. To answer these questions, in this study, we co-cultured NSCs with VECs in order to imitate the situation in vivo, in which NSCs are associated with the endothelium, and treated the single-cultured NSCs and the co-cultured NSCs with safrole oxide. The results showed that safrole oxide (10-100 microg/mL) had no effects on NSC growth. Based on these results, we treated the co-culture system with this small molecule. The results showed that the NSCs differentiation, into neurons and gliacytes was induced by VECs untreated with safrole oxide. But in the co-culture system treated with safrole oxide, the NSCs underwent apoptosis. The data suggested that when VEC apoptosis occurred in the co-culture system, the NSC survival and differentiation could not be maintained, and NSCs died by apoptosis. Our finding provided a useful tool for investigating the effect of apoptosis in vascular endothelial cells on neural stem cell survival and differentiation in vitro.