Bone Marrow Stromal Cells Regulate The Proliferation, Survival And Differentiation Of Neural Stem Cells By CINC-3

Objectives To investigate the effect of non-contact and co-culture of bone marrow stromal cells(BMSCs)and neural stem cells(NSCs)on the secretion of cytokine-induced neutmphil chemoattractant-3(CINC-3)by BMSCs,and to study the effects of CINC-3 on the survival,proliferation and differentiation of neonatal rat hippocampal NSCs.Methods 1 The effect of co-culture of BMSCs and NSCs on the secretion of CINC-3 was divided into NSCs group,BMSCs group,contact co-culture group and non-contact coculture group.The secretion of CINC-3 was detected by ELISA kit;2 The effect of CINC-3 on the survival and proliferation of NSCs is divided into control group and CINC-3group.The cell viability was detected by MTS.Observe cell survival and proliferation by cell growth curve.The CINC-3 concentration with the best NSCs cell viability in MTS assay was used for the next experiment.The expression of Nestin in NSCs was detected by immunofluorescence staining and RT-PCR to determine the effect of CINC-3 on the proliferation of NSCs;3 The effect of CINC-3 on the differentiation of NSCs was divided into two groups: control group and CINC-3 group.The Nestin,a neuron marker,microtubule associated protein2(MAP-2),glial fibrillary acidic protein(GFAP)and oligodendrocyte marker(O4)were determined by immunofluorescence staining and RTPCR.In order to clarify the effect of CINC-3 on the differentiation of NSCs;4 The data were processed by SPSS 21.0 software package.The experimental results were expressed by x ±s.T-test was used for comparison between two groups,One-way ANOVA analysis and SNK for pairwise comparison,P<0.05 was considered statistically significant.Results 1 ELISA results showed that CINC-3 secretion was increased in the contact coculture group,non-contact co-culture group and BMSCs group compared with NSCs group,and the differences were statistically significant(P<0.05);the amount of CINC-3 secreted by the contact co-culture group was higher than that of the non-contact co-culture group,the difference was statistically significant(P<0.05);there was no significant difference between non-contact co-culture group and BMSCs group(P>0.05).Among all the groups,the CINC-3 secretion was the highest in the contact co-culture group.2 The MTS results showed that the cell viability of 1ng/ml CINC-3 group was not significantly different from that of the control group(P>0.05),the cell viability of 5ng/ml,10ng/ml,20ng/ml CINC-3groups were higher than that of the control group,the difference were statistically significant(P<0.05),among which 10 ng/ml CINC-3 group was the best,there was no significant difference between 20 ng/ml CINC-3 group and 10 ng/ml CINC-3(P>0.05).3To detect the effect of CINC-3 on the proliferation of NSCs: the results of immunofluorescence double detection showed that the percentage of Nestin positive cells in the CINC-3 group was significantly higher than that in the control group,and the number of Nestin positive cells in the CINC-3 group was significantly higher than that in the control group(P<0.05).The RT-PCR showed that the expression level of Nestin in CINC-3 group increased significantly,and the expression level was significantly higher compared with control group,and the difference was statistically significant(P<0.05).4To detect the effect of CINC-3 on the differentiation of NSCs: the immunofluorescence staining showed that GFAP and MAP-2 and O4 showed a significant increase in the expression level It was significantly higher than the control group,and the difference was statistically significant(P<0.05).The results of RT-PCR showed that the CINC-3 group still expressed Nestin,but the expression level was significantly decreased,and the expression level was significantly lower than that of the control group(P<0.05),while the m RNA expression levels of MAP-2 and GFAP were significantly increased,and the expression level was significantly higher than that of the control group(P<0.05).Conclusions 1 Co-culture of BMSCs with NSCs can promote the secretion of CINC-3 by BMSCs;2 CINC-3 can promote the survival and proliferation of hippocampal-derived NSCs;3 CINC-3 can promote the differentiation of hippocampal NSCs into astrocytes and neurons.Figure 9;Table 4;Reference 148...