慢病毒载体介导增强绿色荧光蛋白感染人脂肪间充质干细胞及其成脂分化的实验研究

Experimental study of h ADSCs infected by lentiviral vector with EGFP and adipogenic differentiation

目的探讨慢病毒载体介导增强绿色荧光蛋白(EGFP)感染人脂肪间充质干细胞(h ADSCs)的最佳条件与其成脂分化能力。方法以不同感染复数(MOI)感染h ADSCs8h,在荧光倒置显微镜下观察感染效果;流式细胞仪检测感染率;四甲基偶氮唑蓝(MTT)法检测感染前后h ADSCs的增殖情况;成脂诱导14 d后油红O染色,Image-Pro Plus图像分析软件比较h ADSCs在感染前后的成脂能力。结果以MOI值1、10、25、50、75、100、150感染h ADSCs时,其感染率依次为0.24%、13.48%、40.33%、89.27%、94.52%、98.49%和99.11%。以MOI值50感染h ADSCs,比较感染前后细胞的增殖与成脂能力,差异无统计学意义(P>0.05)。结论慢病毒感染h ADSCs的最佳MOI值为50。

Objective To explore the best conditions of lentiviral vector infecting human adipose tissue-derived stromal cells （hADSCs） and its adipogenic differentiation ability before and after infected.Methods After adipose tissue-derived stromal cells had been infected for 8 hours by lentiviral with EGFP at different MOI,effect of infection was observed with inverted fluorescent microscope,and the infection rate was analyzed by FCM （flow cytometry）.The proliferation abilities of hADSCs were compared before and after infected by MTT.After induction of 14 days and stained by Oil red O,the adipogenic ability was analyzed for infection and no infection by The Image-Pro Plus Image analysis software.Results The efficiency of infection at MOI 1,10,25,50,75,100 and 150 was 0.24％,13.48％,40.33％,89.27％,94.52％,98.49％ and 99.11％,the proliferation and adipogenic ability of hADSCs infected had no significant effect at MOI 50,when compared with hADSCs without infection.Conclusion MOI 50 was optimal conditions for lentiviral vector infecting hADSCs.