中国组织工程研究Issue(41)：6603-6608,6.DOI:10.3969/j.issn.2095-4344.2014.41.008
Let-7d慢病毒载体体外诱导大鼠骨髓间充质干细胞分化为神经细胞
Let-7d lentiviral vector induces the differentiation of rat bone marrow mesenchymal stem cells into neurons in vitro
摘要
Abstract
BACKGROUND:MicroRNA plays an important role in the process of growth and aging of living body. To know the role of let-7d in inducing bone marrow mesenchymal stem celldifferentiation into neurons can promote the stem celltransplantation. OBJECTIVE:To investigate the role of let-7d in inducing bone marrow mesenchymal stem celldifferentiation into neurons. METHODS:(1) The lentiviral vector of let-7d was constructed and transfected into rat bone marrow mesenchymal stem cells. The cells were divided into non-transfected group, negative control group (transfected with FU-RNAi-NC-LV), transfected enhancement group (transfected with let-7d-LV), transfected inhibition group ( transfected with let-7d-inhibition-LV). (2) Rat bone marrow mesenchymal stem cells were treated with fasudil as an inducer for triggering the cells to differentiate into neurons. The expression of neuron-specific markers, neuron-specific enolase and microtubule-associated protein 2, were measured by immunocytochemical method. The mRNA expression of microtubule-associated protein 2 was detected by RT-PCR. The viability of bone marrow mesenchymal stem cells was determined by MTT method. RESULTS AND CONCLUSION:Under inverted fluorescence microscope, the cells were successful y transfected with let-7d. Fasudil induced bone marrow mesenchymal stem cells to differentiate into neurons. The transfection efficiency and expression levels of neuron-specific enolase and microtubule-associated protein 2 in transfected enhancement group were higher than those in the negative control group (P<0.05);while in the inhibition group, they were lower than those in the negative control group (P<0.05). These findings indicate that let-7d can promote the differentiation of bone marrow mesenchymal stem cells into neurons induced by fasudil, and by control ing the expression of let-7d we can influence the differentiation efficiency from bone marrow mesenchymal stem cells to neurons.关键词
干细胞/骨髓干细胞/骨髓间充质干细胞/let-7d/法舒地尔/神经元/诱导分化/慢病毒/国家自然科学基金Key words
bone marrow/mesenchymal stem cells/lentivirus/neurons分类
医药卫生引用本文复制引用
徐小隔,张静,龚哲,赵绍云,何霞,王天舒,焦淑洁,滕军放,贾延劼..Let-7d慢病毒载体体外诱导大鼠骨髓间充质干细胞分化为神经细胞[J].中国组织工程研究,2014,(41):6603-6608,6.基金项目
国家自然科学基金项目(骨髓间质干细胞神经命运决定中的自噬现象及其mTOR信号通路的作用/81071114、Lin28/Let-7调控的骨髓间充质干细胞移植治疗阿尔茨海默病的实验研究/81371385) （骨髓间质干细胞神经命运决定中的自噬现象及其mTOR信号通路的作用/81071114、Lin28/Let-7调控的骨髓间充质干细胞移植治疗阿尔茨海默病的实验研究/81371385）
