阳离子多聚物jetPEI^(TM)对原代培养的猪血管平滑肌细胞的转染效率研究

Study of transfection efficiency of cationic polymer PEI to primary culture vascular smooth muscle cells of porcine

目的研究转染试剂jetPEITM对原代培养的猪血管平滑肌细胞的转染效率。方法原代培养小型猪胸主动脉血管平滑肌细胞,体外合成双链寡核苷酸,jetPEITM与寡核苷酸按不同的氨基与磷酸基的比例(N/P)混合转染细胞,采用流式细胞仪和共聚焦荧光扫描仪检测jetPEITM对寡核苷酸的转染效率及细胞内定位情况,MTT法检测jetPEITM的细胞毒性作用。结果原代培养的猪血管平滑肌细胞jetPEITM转染率在N/P=10时可高达(90.89±2.23)%,转染底物主要分布在细胞浆内,核内转染效率极低,在N/P比值在5～10之间时,细胞毒性作用不高于20.00%。结论在原代培养的猪的血管平滑肌细胞,jetPEITM可以高效运送寡核苷酸进入细胞浆内,细胞毒性作用低。

[Objective] To study the transfection efficiency of the cationic polymer jetPEI to primary culture vascular smooth muscle cells of the porcine, [Methods] We cultured vascular smooth muscle cells of the pig and synthesized oligonucleotides in vitro, After transfected at different ratio of N/P, we applied FACS and laser confocal microscopy to detect the transfection efficiency and the location of the transfection substances in cell, Then we also used MTr to evaluate the cytotoxin action of the agent, [Results] We found that when N/P=10, the transfection efficiency of jetPEI^TM to primary culture smooth muscle cell was about （90.89±2.23）%, and the transfection substances mainly located at cytoplasm with little in nucleus. When the ratio of N/P ranged from 5 to 10, cytotoxin action was less than 20.00%. [Conclusion] jetPElTM is a high efficient transfection agent to deliver the oligonucleotides into cytoplasm in the primary culture smooth muscle cells and the cytotoxin action is low.