转染试剂JetPEI和Lipofectamine Plus对NIH3T3转染效果及毒性的比较和评价  

Comparison and evaluation of transfection efficiency and toxicity of two transfection reagents:JetPEI and Lipofectamine Plus in NIH3T3

作　　者：张世新[1] 杨康[1] 廖克龙[1] 王海东[1] 吴蔚[1] 刘鸿翔[1] 张现普[1] 吴涛[1] 

机构地区：[1]第三军医大学西南医院胸心外科,重庆400038

出　　处：《重庆医学》2011年第12期1164-1166,共3页Chongqing medicine

摘　　要：目的比较及评价两种转染试剂JetPEI和Lipofectamine Plus对小鼠胚胎成纤维细胞(NI H3T3)的转染效果及毒性。方法分别用阳离子聚合物JetPEI和阳离子脂质体Lipofectamine Plus将表达绿色荧光蛋白(GFP)的质粒转染NI H3T3,24h后,用流式细胞仪检测转染效率;分别用JetPEI和Lipofectamine Plus将75 nM24 bp的双链寡脱氧核苷酸(ODN)转染NI H3T3,12 h后,用激光共聚焦显微镜观察ODN在细胞内的分布;分别将转染相同量DNA的JetPEI和Lipofectamine Plus加入细胞培养基中,48 h后,用WST-8试剂检测其对细胞的毒性。结果对于大分子质粒,JetPEI的转染效率更高;对于小分子ODN,JetPEI包裹ODN后形成的颗粒更小,更易于进入细胞核内;转染相同量的DNA,JetPEI对细胞的毒性更小。结论对于转染NI H3T3,JetPEI是一种更理想的转染试剂。Objective To comparison and evaluation of transfection efficiency and toxicity of two transfection reagents：JetPEI and Lipofectamine Plus in NIH3T3. Methods Vector expressing GFP was transfected into NIH3T3 cells mediated by JetPEI or Lipofectamine Plus. 24 houes later, transfection rate was detected by flow cytometry. 75nM Oligodeoxynucleotide（ODN）was trans fected into NIH3T3 cells mediated by JetPEI or Lipofectamine Plus： 12 hours later, the distribution of ODN in the cells was investigated by laser scanning confocal fluorescence microscope. NIH3T3 cells were transfected with JetPEI and Lipofectamine Plus respectively. 48 hours later,the toxicity of transfection reagent to cells was examined by WST-8. Results JetPEI had higher transfection rate than Lipofectamine Plus when NIH3T3 cells were transfected with macromolecule vector. To micromolecule ODN, It seams that transfection rate was no difference between JetPEI and Lipofectamine Plus, but the DNA particles that JetPEI compacted were smaller than Lipofectamine Plus compacted,so it was easier for ODN to enter the cell nucleus. JetPEI had lower cell toxicity than Lipofectamine Plus when NIH3T3 cells were transfected with 200nM ODN. Conclusion JetPEI has higher transfection rate and lower toxicity than Lipofectamine Plus to NIH3T3.

关 键 词：转染 成纤维细胞 毒性 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]