This study aimed to determine the effects of the controlled release of brain-derived neurotrophic factor (BDNF) from collagen gel on rat neural stem cells (NSCs). With two groups of daily addition of BDNF and collagen gel without BDNF as controls, BDNF was tested using ELISA at different time points. In the BDNF-collagen gel group, BDNF was steadily released from gels for 10 days. The cell viability test and the bromodeoxyuridine incorporation assay showed that the BDNF-collagen gel supported the survival and proliferation of NSCs. Compared with controls, the length of processes was markedly longer and the differentiation percentage from NSCs into neurons was much higher in the BDNF-collagen gel group (P<0.05). These findings suggest that BDNF-collagen gel can significantly reduce the amount of BDNF required for the culture of NSCs and increase the differentiation percentage from NSCs into neurons.