过表达BDNF基因重组慢病毒转染MSCs的实验研究

Over expression of BDNF in MSCs by transfection of recombinant lentiviral vector

目的观察BDNF基因重组慢病毒修饰MSCs过表达BDNF的情况。方法构建携带BDNF基因的慢病毒载体;分离培养大鼠MSCs;将基因重组慢病毒转染MSCs;RT-PCR、Western blot检测各组BDNF基因及蛋白表达水平。结果绿色荧光试验证实BDNF基因重组慢病毒转染MSCs成功;RT-PCR、Western blot验证MSCs-EGFP-BDNF组BDNF基因及蛋白表达明显高于MSCs组及MSCs-EGFP组,MSCs组与MSCs-EGFP组比较差异无统计学意义。结论 BDNF基因重组慢病毒修饰的MSCs基因及蛋白表达均增高,MSCs成功过表达BDNF。

Objective To observe over expression of BDNF in MSCS modified by recombinant lentivirus. Methods We es- tablished the lentiviral vector carrying BDNF gene; MSCs from rat brain was cultured; the lentiviral vector carrying BDNF was transfected into MSCs; RT-PCR and Western blot were used to detect the expression of BDNF gene and protein in each group. Re- suits The transfection of BDNF recombinant lentiviral vector into MSCs was verified through green fluorescent assa. RT-PCR and Western blot showed that the expression of BDNF gene and protein in MSCs-EGFP-BDNF group was higher than that in MSCs group and MSCs-EGFP group. There was no statistical difference between the MSCs group and the MSCs-EGFP group. Conclusion BDNF gene and protein were successfully over expressed by transfection of BDNF recombinant lentiviral vector into MSCs.