Interference Hes1 Gene To Bone Marrow Stromal Cells To The Cholinergic Neurons In Vitro Pre-

Objective: To explore the expression of Mash1 after the inhibition of Hes1 gene inBMSCs.Methods: BMSCs isolated from bone marrow were cultured in vitro. The morphologicfeature and growth status of BMSCs and differentiated BMSCs were observed by invertmicroscope. The expression of Mash1 was analyzed by real-time quantitative PCR afterthe inhibition of Hes1 gene in BMSCs.Results: The concentration of Hes1 siRNA were selected for 200nM, 100nM, 50nMand 25nM. The expression of Hes1 was low-decreased in comparison to control group.The level of Hes1 at the concentration of 200nM in interfered BMSCs was 0.036-fold ofthat in control BMSCs (P<0.05),and the level of Hes1 at the concentration of 50nM ininterfered BMSCs was 0.076-fold of that in control BMSCs (P<0.05), and they werebetter than the others. The expression of Mash1 gene was increased after the inhibitionof Hes1 gene with a dose-dependent effect.Conclusion: Hes1 can down-regulate the expression of Mash1 gene in BMSCs. Objective: To discuss the mechanism of cholinergic neurons differentiate induced frombone marrow stromal cells in vitro by analysising the impact of interfering expression ofthe gene Hes1 to the expression of the gene Sept-9 and Stmn1.Methods: The third generation of bone marrow stromal cells,seeded in polylysinecoated6-well culture plate,was induced to cholinergic neurons differentiation byadding the inducible factor RA, SHH and bFGF in the culture medium. To analysis theexpression of the gene Sept-9 and Stmn1 of interfering the gene Hes1 in the process ofBMSCs induced to cholinergic neurons differentiation by real time and quantitativePCR.Results: The expression of the gene Hes1 increased, after alone induced; Theexpression of the gene Hes1 increased significantly in the induction and differentiationgroup after interfered. The expression of the gene Sept-9 increased, after siRNA (50nM)interfered. There was no difference the expression of the gene Sept-9 compared with thecontrol group, when the BMSCs of P3 was alone induced in vitro. The expression of thegene Sept-9 increased significantly, when the BMSCs of P3 was induced after the geneHes1 was interfered. The expression of the gene Stmn1 increased significantly, whenthe gene Hes1 was interfered by siRNA (50nM); The expression of the gene Stmn1increased slightly, when the BMSCs of P3 was alone induced; The expression of thegene Stmn1 increased significantly, when the BMSCs of P3 was induced after the geneHes1 was interfered, and the increased range was among the interfered group andinduced group .Conclusion: It was very important for the expression of the gene Sept-9, Stmn1 tointerfer the expression level of the gene Hes1.