中国实用神经疾病杂志Issue(23)：1-4,4.
Let-7b 慢病毒载体构建及对大鼠骨髓间充质干细胞诱导分化为神经细胞的影响
Effect of lentiviral vector of Let-7b on differentiation of rat bone marrow mesenchymal stem cells into neurons
张静 1徐小隔 1龚哲 1赵绍云 1何霞 1王天舒 1焦淑洁 1滕军放 1贾延劼1
作者信息
- 1. 郑州大学第一附属医院神经内科 郑州 450052
- 折叠
摘要
Abstract
Objective To investigate the role of Let‐7b in inducing bone marrow mesenchymal stem cell(MSCs) differenti‐ation into neurons.Methods The lentiviral vector of Let‐7b up (LV‐rno‐Let‐7b‐up) and inhibition (LV‐rno‐Let‐7b‐inhibition) was constructed and transfected into rat MSCs.The cells were divided into non‐transfected group ,transfected group 1(trans‐fected with LV‐rno‐Let‐7b‐up) ,transfected group 2 (transfected with LV‐rno‐Let‐7b‐inhibition) and negative control group (transfected with empty virus ).MSCs were treated with Fashudier as an inducer for triggering the cells to differentiate into neurons.The fluorescence expressed by transfected MSCs were observed under inverted fluorescence microscope.The mRNA expression of microtublin‐associated protein 2 (MAP‐2) was detected by RT‐PCR.The expression of neuron‐specific markers , neuron‐specific enolase(NSE) and MAP‐2 were measured by immunocytochemical method.The viability of MSCs was deter‐mined by MTT method.Results In LV‐rno‐Let‐7b‐up the best transfection efficiency(up to 92.1% ± 4.3% ) and survival rate appeared when multiply of infection (MOI) was 10 and on 3rd day.In LV‐rno‐Let‐7b‐inhibition the best transfection efficiency (up to 90.3% ± 4.2% ) and survival rate appeared when MOI was 20 and on 5th day.Fashudier induced MSCs to differentiate into neurons and the best efficiency of the induction was observed in transfected group 1.The expression levels of NSE and MAP‐2 in transfected cells were higher than those in the cells of other group ( P<0.05).Conclusion LV‐rno‐Let‐7b‐up has high transfection efficiency in rat MSCs.Higher differentiation rate from rat MSCs to neurons is induced by Fashudier after the cells are transfected with LV‐rno‐Let‐7b‐up.关键词
Let-7b/骨髓间充质干细胞/神经元Key words
Let-7b/Marrow mesenchymal stem cells/Neurons分类
医药卫生引用本文复制引用
张静,徐小隔,龚哲,赵绍云,何霞,王天舒,焦淑洁,滕军放,贾延劼..Let-7b 慢病毒载体构建及对大鼠骨髓间充质干细胞诱导分化为神经细胞的影响[J].中国实用神经疾病杂志,2014,(23):1-4,4.
