Study of anti-apoptotic effect of erythropoietin on the cultured NSCs from brain cortex of embryonic rats in vitro

Objective To explore the effects of EPO on the apoptosis of the neural stem cells(NSCs) in vitro from brain cortex of embryonic Sprague-Dawley rats.Methods The NSCs were isolated and cultured in serum-free suspension,then cultured by differentiating suspension.Nestin was used to detect the NSCs.Microtubule-associated protein 2(MAP2) and glial fibrillary acid protein(GFAP) was used to detect the differentiation of NSCs by immunofluorescent cytochemistry.EPO was added to the serum-free suspension.According to the final EPO concentration,the cells were divided into 4 experimental groups,they were 0.5,5,50,500U/ml,in addition the control group was without EPO.Caspase-3 was measured to analyze the apoptosis of NSCs by immunofluorescent cytochemistry.Results The results showed that separated cells from brain cortex could proliferate and subculture in vitro.Nestin-positive cells were detected in neural stem spheres.MAP2-and GFAP-positive cells were detected in serum with 10% FBS.Caspase-3-positive NSCs in EPO supplementation groups(5U/ml) were decreased in neural stem spheres compared with the control group.Conclusion Our results indicate that the separated cells from brain cortex of embryonic rats are multipotent and have the ability of self-renew and differentiation,which suggest that it may be a good cell culture model for the study of NSCs.The present results suggest that EPO can decrease the apoptosis of the NSCs in vitro.