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

作　　者：吴婷[1,2,3,4] 吴江 金书欣[3,4] 席晔斌[3] 钮晓音 陈广洁 WU Ting1,2,3,4 ,WU Jiang2 , JIN Shu-xin3,4, XI Ye-bin3 , NIU Xiao-yin3,4 , CHENG Guang-jie3(1. Renji Hospital Shanghai Jiaotong University School of Medicine, Embryo Laboratory, Department of Reproductive Medicine, Shanghai 200135, China; 2. Renji Hospital Shanghai Jiaotong University School of Medicine, Department of Blood Transfusion, Shanghai 200127, China; 3. Department of Immunology and Microbiology, Shanghai Jiaotong University School of Medicine, Shanghai 200025, China ; 4. Shanghai Institute of Immunology, Shanghai 200025, Chin)

机构地区：[1]上海交通大学医学院附属仁济医院生殖医学科胚胎实验室,上海200127 [2]上海交通大学医学院附属仁济医院输血科,上海200127 [3]上海交通大学医学院免疫学与微生物学系,上海200025 [4]上海市免疫学研究所,上海200025

出　　处：《现代免疫学》2018年第2期89-94,共6页Current Immunology

基　　金：国家自然科学基金(81771731)

摘　　要：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.

关 键 词：阳离子脂质体 转染效率 贴壁细胞 悬浮细胞 原代细胞 

分 类 号：R392.11[医药卫生—免疫学]