Ebadi Reihane, Kordi-Tamandani Dor Mohammad, Ghaedi Kamran, Nasr-Esfahani Mohammad Hossein
Department of Biology, University of Sistan and Baluchestan, P.O. Box 98155-411, Zahedan, Iran.
Department of Biology, Faculty of Sciences, University of Isfahan, Isfahan, Iran.
Mol Biol Rep. 2018 Dec;45(6):2377-2391. doi: 10.1007/s11033-018-4404-4. Epub 2018 Oct 10.
Neural cells derived from embryonic stem cells (ESCs) have potential usefulness for the treatment of neurodegenerative disorders. Modulation of intrinsic growth factors expression such as neurotrophins and their respective receptors by these cells is necessary to obtain functional neural cells for transplantation. In present study, we compared neural differentiation potential of two different media, NB + 5%ES-FBS + N2B27 and Ko-DMEM + 5%ES-FBS for conversion of mESC derived neural progenitors (NPs) into mature neural cells with emphasis on effect of the these two media on neurotrophins and their respective receptors expression. Immunofluorescence staining, RT-qPCR and western blot analysis showed that the expression of neuronal specific markers, MAP2 and Tuj-1, in NB + 5%ES-FBS + N2B27 medium was significantly higher than the other medium. Western blot assay revealed that the expression of BDNF and NGF increased significantly in mature neural cells obtained from NB + 5%ES-FBS + N2B27 medium but decreased in neural cells from Ko-DMEM + 5%ES-FBS medium compared to mESCs. TrkB protein was not detectable in mESCs but its expression increased in neural cells obtained from both media although its expression in NB + 5%ES-FBS + N2B27 medium was significantly higher than the other medium. In contrast to TrkB, p75NTR protein was detectable in mESCs and is remained constant in neural cells cultured in NB + 5%ES-FBS + N2B27 medium but decreased significantly in the other medium. In conclusion, our results indicated that NB + 5%ES-FBS + N2B27 medium promoted neural differentiation process of mESCs and caused enhancement of neurotrophins protein expression in addition to their cognate receptors.
源自胚胎干细胞(ESC)的神经细胞在治疗神经退行性疾病方面具有潜在用途。这些细胞对神经营养因子等内在生长因子及其各自受体的表达进行调节,对于获得用于移植的功能性神经细胞是必要的。在本研究中,我们比较了两种不同培养基(NB + 5%ES-FBS + N2B27和Ko-DMEM + 5%ES-FBS)将小鼠胚胎干细胞(mESC)来源的神经祖细胞(NP)转化为成熟神经细胞的神经分化潜能,重点关注这两种培养基对神经营养因子及其各自受体表达的影响。免疫荧光染色、RT-qPCR和蛋白质印迹分析表明,在NB + 5%ES-FBS + N2B27培养基中,神经元特异性标志物MAP2和Tuj-1的表达显著高于另一种培养基。蛋白质印迹分析显示,与mESC相比,从NB + 5%ES-FBS + N2B27培养基获得的成熟神经细胞中BDNF和NGF的表达显著增加,但在Ko-DMEM + 5%ES-FBS培养基的神经细胞中则降低。TrkB蛋白在mESC中无法检测到,但其在两种培养基获得的神经细胞中均有表达,尽管其在NB + 5%ES-FBS + N2B27培养基中的表达显著高于另一种培养基。与TrkB相反,p75NTR蛋白在mESC中可检测到,并且在NB + 5%ES-FBS + N2B27培养基中培养的神经细胞中保持恒定,但在另一种培养基中显著降低。总之,我们的结果表明,NB + 5%ES-FBS + N2B27培养基促进了mESC的神经分化过程,并导致神经营养因子及其同源受体的蛋白质表达增强。