The Expression Of MicroRNA-223 And RhoB On The Spinal Cord-derived Neural Stem Cells Of Embryonic Rats

Objective To obtain, cultivate and authenticate the neural stem cells(NSCs) from rat embryonic spinal cord in vitro for further related research.Methods The spinal cord was dissected out from green fluorescent protein(GFP) transgenicembryonic rats,and enzymatically and mechanically dissociated into single cell suspension, the cells were cultured with serum-free medium containing DMEM/F12(1:1),N2 and B27 supplement,EGF and b FGF. After cultivation and fetal bovine serum-induced proliferation,the neural stem cells were identified by immunofluorescence technique for the expression of Nestin、Neu N(Neuronal nuclei)and GFAP(Glial fibrillary acidic protein).Results The neural cells obtained from rat embryonic spinal cord growth well in serum-free medium in vitro.The immunocytochemistry showed most of the cells had positive Nestin expression. After serum induced differentiation, NSCs also could express positive antigen of GFAP and Neu N.Conclusion The neural stem cells acquired from rat embryonic spinal cord could be successfully separated and culturied, which would lay a root for further research.Objective To detect the expression of Rho B Protein and mi RNA-223 in neural stem cells, meanwhile preliminarily investigate the relationship between them.Methods Neural stem cells from E14、E16、E18 and E20 rat embryos were isolated and cultured respectively;Western Blot and quantitative QPCR analysis were performed to detect the expression of Rho B protein and mi RNA-223 in each group. At the same time, the correlation between mi RNA-223 and Rho B were analyzed by statistical methodsResults The quantitative QPCR analysis showed mi R-223 expressed in all four groups. The expression of mi R-223 is higher in E18 group than other three groups, which showed significant statistical difference. Western Blot results showed Rho B protein expressed in all four groups, and the Rho B expression is higher in E14 and E16 groups, lower in E18 and E20 groups. Spearman correlation analysis showed that there is significant negative correlation(r=-0.875,P<0.001) between the expressions of Rho B protein and mi R-223 in the NSCs of four groups.Conclusions mi R-223 and Rho B protein can steadily express on the Neural stem cells, meanwhile the correlation between them may be statistically negative.