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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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[1] Ma K,Fox L,Shi G,Generation of neural stem cell-like cells from bone marrow-derived human mesenchymal stem cells.Neurol Res, 2011,33(10):1083-1093.
[2] Friedenstein A J, Gorskaja J F, Kulagina N N.Fibroblast precursors in normal and irradiated mouse hematopoietic organs.Exp Hematol, 1976,4(5):267-274.
[3] Polisetti N, Chaitanya V G, Babu P P, et al.Isolation, characterization and differentiation potential of rat bone marrow stromal cells.Neurol India, 2010,58(2):201-208.
[5] Zhu H, Guo Z K, Jiang X X,et al.A protocol for isolation and culture of mesenchymal stem cells from mouse compact bone.Nat Protoc, 2010,5(3):550-60.
[5] Chen B Y, Wang X, Chen L W,et al.Molecular targeting regulation of proliferation and differentiation of the bone marrow-derived mesenchymal stem cells or mesenchymal stromal cells.Curr Drug Targets, 2012,13(4):561-571.
[6] Tao H, Rao R, Ma D D.Cytokine-induced stable neuronal differentiation of human bone marrow mesenchymal stem cells in a serum/feeder cell-free condition.Dev Growth Differ,2005,47(6):423-433.
[7] Rezaei M,Karbalaie K,Tanhaie S,et al.bone morphogenetic protein-4 influences neural differentiation of induced mouse mesenchymal stem cells.Cell Journal,2011,12(4):511-516.
[8] Lai S L, Chien A J, Moon R T.Wnt/Fz signaling and the cytoskeleton: potential roles in tumorigenesis.Cell Res, 2009,19(5):532-545.
[9] Tang M, Miyamoto Y, Huang E J.Multiple roles of beta-catenin in controlling the neurogenic niche for midbrain dopamine neurons.Development, 2009,136(12):2027-2038.
[10] David M D, Cantí C, Herreros J.Wnt-3a and Wnt-3 differently stimulate proliferation and neurogenesis of spinal neural precursors and promote neurite outgrowth by canonical signaling.J Neurosci Res, 2010,88(14):3011-3023.
[11] Wislet-Gendebien S, Hans G, Leprince P, et al.Plasticity of cultured mesenchymal stem cells: switch from nestin-positive to excitable neuron-like phenotype.Stem Cells, 2005,23(3):392-402.
[12] 陈丹丹,付文玉,庄文欣,等.多种细胞因子诱导大鼠间充质干细胞向多巴胺能神经元分化.解剖学杂志,2011,34(5):584-587. Chen D D, Fu W Y, Zhuang W X, et al. Some cytokines induce rat bone marrow mesenchymal stem cells to differentiate into dopaminergic neurons in vitro. Chinese Journal of Anatomy, 2011,34(5):584-587.
[13] Brunet N, Tarabal O, Portero-Otín M, et al.Survival and death of mature avian motoneurons in organotypic slice culture: trophic requirements for survival and different types of degeneration.J Comp Neurol, 2007,501(5):669-690.
[14] Li Q, Ping P, Jiang H, et al.Nerve conduit filled with GDNF gene-modified Schwann cells enhances regeneration of the peripheral nerve.Microsurgery, 2006,26(2):116-121.
[15] 吴晓娟,魏明发,柴成伟,等.大鼠骨髓间充质干细胞体外向肠神经细胞分化及胶质细胞源神经营养因子表达的变化.中华小儿外科杂志,2010,31(8):607-611. Wu X J, Wei M F, Chai CH W, et al. Differentiation of rat bone marrow stromal cells into enteric neurons and expression change of glial cell line-derived neurotrophic factor. Chinese Journal of Pediatric Surgery, 2010,31(8):607-611.
[16] Tanaka J, Horiike Y, Matsuzaki M, et al.Protein synthesis and neurotrophin-dependent structural plasticity of single dendritic spines.Science, 2008,319(5870):1683-1687.
[17] Yoshii A, Constantine-Paton M.Postsynaptic BDNF-TrkB signaling in synapse maturation, plasticity, and disease.Dev Neurobiol, 2010,70(5):304-322.
[18] Chen B Y, Wang X, Wang Z Y,et al.Brain-derived neurotrophic factor stimulates proliferation and differentiation of neural stem cells, possibly by triggering the Wnt/β-catenin signaling pathway.J Neurosci Res, 2012,91(1):30-41.
[19] Sanalkumar R, Vidyanand S, Lalitha Indulekha C,et al.Neuronal vs. Glial Fate of Embryonic Stem Cell-Derived Neural Progenitors (ES-NPs) is Determined by FGF2/EGF During Proliferation.J Mol Neurosci, 2010,42(1):17-27.
[20] Ohinata Y, Ohta H, Shigeta M, et al.A signaling principle for the specification of the germ cell lineage in mice.Cell, 2009,137(3):571-584. |
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