阳离子脂质体Lipofectamine 3000对贴壁细胞、悬浮细胞和原代细胞转染效率比较的研究

The study on the transfection efficiency of cationic liposome lipofectamine 3000 in adhesive cells,suspension cells and primary cells

为了比较阳离子脂质体Lipofectamine 3000(Lipo3000)在不同细胞类型中的转染效率,课题组分别在贴壁细胞NIH3T3、悬浮细胞EL4和原代细胞CD4+T细胞中,利用Lipo3000将不同浓度的miRNA mimics转染至细胞内,通过流式细胞术检测的方法,比较不同细胞之间以及同一细胞、不同浓度的miRNA mimics组之间转染效率的差异。结果显示,在NIH3T3细胞中,miRNA mimics为100nmol/L时,转染效率最高,可达(41.47±8.10)%;而在EL4和CD4+T细胞中,随着miRNA mimics浓度的增加,转染效率呈上升的趋势,最高分别可达(12.13±1.16)%和(3.80±0.60)%。可见,Lipo3000在贴壁细胞和悬浮细胞的转染效率明显高于原代细胞。

To compare the transfection efficiency of cationic liposome lipofectamine3000（Lipo3000）in different cell types,miRNA mimics with different concentrations were transfected into NIH3 T3,EL4 and CD4^＋T cells respectively by Lipo3000.The differences in transfection efficiency were compared by FACS among different types of cells and among different concentrations of miRNA mimics groups in the same types of cells.The results indicated that the highest transfection efficiency in NIH3 T3 would be obtained when transfected with 100 nmol/L miRNA mimics by Lipo3000.For EL4 and CD4^＋T cells,the efficiency of transfection was improved with the increase of miRNA mimics concentration and could reach up to（12.13±1.16）% and（3.80±0.60%）respectively.It could be concluded that the transfection efficiency of Lipofectamine 3000 for adherent cells and suspension cells was significantly higher than that in primary cells.