Introduction: After primary brain injury, secondary mechanisms are activated and lead to releasing of various supportive and neurotrophic factors in injured tissues. The effect of the new environment on neurogenesis, proliferation, and survival of stem cells needs more investigations. The aim of the present study was to evaluate the effect of injured brain extract on proliferation of embryonic rat neural stem cells (NSCs). Materials and Methods: NSCs were isolated from ganglionic eminences of embryonic rat then cultured as neurospheres. Next, cells were seeded in PuraMatrix scaffold as 3-dimension culture. Based on the medium content, cells were divided into 4 groups: with growth factor, without growth factor, without growth factor + intact brain extract, and without growth factor + injured brain extract. Proliferation assay was done by evaluation of the DiI labeled cells and the survival assay was carried out by MTS test 10 days later. For preparation of the injured brain extract, a rat brain injury model was utilized and the extract was collected 48 hours after brain injury. Results: The results showed that NSCs derived ganglionic eminence of embryonic rat had high proliferation ability. DiI-positive cells and MTS test showed a higher tendency of the proliferation and survival of NSCs in the without growth factor + injured brain extract and growth factor groups compared to the without growth factor and without growth factor + intact brain extract groups. Conclusion: Our results indicated a possible positive impact of injured brain extract on survival and proliferation of rat embryonic neural stem cells. Further studies are needed to investigate our preliminary findings in details.