β-catenin慢病毒载体转染大鼠骨髓间充质干细胞的实验研究

Experimental studies on beta-catenin lentiviral vector transfection of rat bone marrow mesenchymal stem cells

【目的】构建β—catenin慢病毒载体,转染骨髓间充质干细胞并予以鉴定。【方法】β—catenin曼病毒载体转染骨髓间充质干细胞,在倒置荧光显微镜下观察转染效率,并进行RNA提取及PCR扩增,检测目的基因表达情况,进行CCK—8实验检测转染后细胞的增殖活性。【结果】当感染复数为25时,感染后96 h观察干细胞的感染效率大约为80%。PCR检测到β—eatenin的mRNA在骨髓间充质干细胞中过表达。慢病毒对细胞的增殖有一定的抑制作用,但β—catenin对细胞增殖有促进作用。【结论】β—catenin慢病毒载体成功转染大鼠骨髓间充质干细胞,为进一步研究β—catenin在MSCs中的作用奠定了基础。

【Objective】To construct β-catenin lentiviral vector,and transfected to rat bone marrow mesenchymal stell cells（BMSCs）and observe its expression in these cells.【Methods】Infection titer was monitored by green eGFP（enhanced green fluorescent protein）,the expression of eGFP were observed under inverted fluorescence microscope and the expression of bale-catenin were observed by PCR amplification.Proliferation activity were tested by CCK-8 test.【Result】 When the multiplicity of infection（MOI） was 25,the infection efficiency of bate- catenin- lentivirus in rat BMSCs was nearly 80%after infection for 96 h and around after 3 weeks of continuous.The over expression of bate-catenin mRNA were detected by PCR amplification in rat BMSCs.Lentivirus could inhibit cell proliferation but B-catenin could promote cell proliferation.【Conclusions】The succesive transfection of β-catenin in the BMSCs by B-catenin lentivirus vector laid the fundation of further study the function of β-catenin in BMSCs.