Objective To observe the effect of the conditioned medium obtained from the coculture of astrocytes and β-amyloid (Aβ1-40)-induced apoptotic PC12 cells on the neuronal differentiation efficiency of neural stem cells (NSCs), and investigate the possible involvement of the neurotrophins proteins including brain-derived neurotrophic factor (BDNF), nerve growth factor (NGF)and neurotrophin-3 (NT-3) in the induced differentiation of the NSCs. Methods PCI2 cells wereinduced by Aβ1-40 for different time lengths (0, 4, 6, 12, and 24 h), and after detection of the apoptotic rates using flow cytometry, the cells were cocultured with astrocytes for 2 days. The astrocyte-conditioned medium (ACM), after analysis of the neurotrophins proteins using erLzyme-linked immunosorbent assay (ELISA), was mixed with DMEM/F12 medium at the proportion of 1:3 to induce the differentiation of the NSCs. The cell differentiation was identified by laser confocal microscopy, NSE immunofluorescent labeling, and neuronal counting. Results Flow cytometry showed that induction by Aβ1-40 for 6 h resulted in the highest apoptosis rate of PC12 cells (P<0.05). BDNF content in the ACM derived from the coculture of astrocytes and the PC12 cells induced for 6 h was significantly increased, and the NSCs induced in this ACM showed the highest neuronal differentiation rates, showing significant difference from those induced by other ACMs (P<0.05). Conclusion The ACM derived from the coculture of Aβ1-40-induced PC12 cells and astrocytes can increase the neuronal differentiation rates of NSCs in vitro, and BDNF in the ACM may play a role in this process.