携载增强型绿色荧光蛋白基因慢病毒标记小鼠脐带间充质干细胞示踪方法

目的 探讨慢病毒载体介导增强型绿色荧光蛋白(enhanced green fluorecent protein,eGFP)转染小鼠脐带间充质干细胞(umbilical cord mesenchymal stem cells,UCMSCs)的最佳感染复数(multiplicity of infection,MOI)。方法 通过组织贴壁法得到mUCMSCs;用生长增殖曲线、成骨、成软骨及成脂分化进行鉴定;以MOI为50、100、150、200携载eGFP的慢病毒载体介导转染mUCMSC,倒置显微镜下观察荧光表达,确定荧光表达率最高的MOI组。结果 eGFP慢病毒以MOI=150转染mUCMSC,效率较高,可直接用于体内示踪。结论 用小鼠脐带可以分离出mUCMSCs,MOI=150时携载eGFP的慢病毒传染mUCMSCs效果最好。

The method of lentiviral vector mediated enhanced green fluorescent proteintransfected into umbilical cord stem cells for labeling and tracing

Objective To investigate the optimal multiplicity of infection (MOI) of lentiviral vector-mediated enhanced green fluorescent protein (eGFP) transfected mouse umbilical cord mesenchymal stem cells (UCMSCs).Methods MUCMSCs was obtained by tissue adherent on the bottom of the culturing bottle.Growth curve,the tests of osteogenic,chondrogenic and lipogenic differentiation were used to identify them.MUCMSCs was transfected with eGFP-carrying lentiviral vectors with MOI of 50,100,150 and 200,and the fluorescence expression was observed under an inverted microscope to determine the best MOI.Results EGFP lentivirus was transfected with mUCMSCs with MOI=150.Conclusion mUCMSC can be isolated from mouse umbilical cord.The effect of lentivirus transfected with eGFP on mUCMSC is best when MOI=150.