Experimental Study On The Effect Of EPO/EPOR Signaling Pathway On The Differentiation Of Neural Stem Cells In Adult Spinal Cord

Objective Adult SD rat spinal cord cells were taken from the primary culture, and the third generation NSCs was taken as the research object, and the recombinant human erythropoietin(rh EPO) was added to the cells in vitro,we aim to study the influence of the erythropoietin(EPO) and erythropoietin receptor(EPOR) signaling pathway on the differentiation of adult rat spinal cord neural stem cells(NSCs).Methods(1)The experiment object: Adult SD rat spinal cord cells were cultured in DMEM/F12 medium and the 3rd generation of the NSCs was chosen as the experimental object(2)Immunofluorescence staining of Brd U and Nestin was performed on cultured adult spinal cord NSCs in order to identify the NSCs.(3)in the experimental group the recombinant human erythropoietin(rh EPO) was further added into the culture medium, meanwhile in a control group only using saline with an equal volume(4)The expression of EPOR protein in NSCs was detected by immunofluorescence staining method(5)EPOR protein expression was detected by Western blot method in each group using(6)By using immunofluorescence staining,we observed the differentiation of NSCsResults(1)After 7 d culture in the microscopically,neurospheres form is the identification of the adult NSCs(2)The results of immunofluorescence staining showed that EPOR expression can be measured in both NSCs of the experimental group and control group,(3)and the experimental group EPOR protein maintained at a higher level than that of the control group.(4)Western blot results showed that the expression level of EPOR protein in the experimental group was higher than that in the control group, the difference was statistically significant(P<0.01).(5)Immunofluorescence staining results of differentiation showed the NSCs of experimental group can differentiate into neurons, whose differentiate proportion was more than the control group, the difference was significant(P<0.01).Conclusion rh EPO can activate EPO/EPOR signaling pathway, thereby enhancing the differentiate ability of adult neural stem cells of spinal cord into neurons.