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Wnt/β-catenin Signaling Pathway In Neuronal Differentiation of Rat BMSCs |
HE Ding-wen1,2, YIN Ming1, WU Ya-hua3, ZHOU Rong-ping1, WEI Qiang-qiang1,2, YIN Chang-chang 3 |
1. The Second Affiliated Hospital Of NanChang University,Nanchang 330006, China; 2. Nanchang University Graduate School of Medicine,Nanchang 330006, China 3 JiuJiang University,Jiujiang 332000, China |
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Abstract Objective To study the role of Wnt / β-catenin signaling pathway in neural differentiation of rat BMSCs, Explore possible signal molecular mechanism in application of EGF and bFGF induce BMSCs to neuronal differentiation.Methods BMSCs were harvested by the whole bone marrow adherence method, BMSCs at passage 3 were respectively treated with 20ng/ml basic fibroblast growth factor (bFGF) and 20ng/ml epidermal growth factor (EGF) in containing 10ml / L (1%) fetal bovine serum (FBS) DMDM/F-12 medium. Then the cellular morphology was observed under inverted phase contrast microscope.At 7days after induction, Immunohistochemical staining was used to detect neuron specific enolase(NSE) and collagen fibre acidic protein(GFAP),and RT-PCR to detect expression of β-catenin、BDNF、GDNF、nestin genes.Results At 7days after induction, BMSCs in group bFGF and EGF+bFGF become round, extend multiple distinct processes around and have connections among some processes,but group EGF and control did not change significantly. Immunohistochemical staining show GFAP positive rate higher than NSE in group EGF,but in group bFGF and EGF+bFGF NSE positive rate higher than GFAP. NSE and GFAP positive rate is highest in group EGF+bFGF, group bFGF is followed, compare With blank group and group EGF,the differences were statistically significant (P<0.0042).RT-PCR show mRNA expression of nestin in group bFGF and EGF+bFGF were significantly higher than other groups(P<0.05); β-catenin、BDNF in group EGF、bFGF and EGF+bFGF were significantly higher than blank group and differences among these three groups were significant (P<0.05). GDNF in group bFGF and EGF+bFGF were significantly higher than other groups and group bFGF was significantly higher than group EGF+bFGF (P<0.05). Conclusion bFGF and EGF are able to induce differentiation of BMSCs into neural cells, the genes of β-catenin、BDNF and GDNF increase expression during differentiation. Wnt / β-catenin signaling pathway may play a important role in neural differentiation of rat BMSCs.
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Received: 11 October 2012
Published: 25 March 2013
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