摘要
目的探索核转染技术转染人外周血淋巴细胞的转染效率。方法利用核转染方法将携带增强型绿色荧光蛋白基因的质粒转染人外周血淋巴细胞,24小时后经流式细胞仪检测绿色荧光蛋白EGFP表达水平,确定转染效率。结果未经任何刺激的外周血淋巴细胞转染效率为8.7%,经抗CD3抗体OKT3和抗CD28抗体共同刺激后的外周血淋巴细胞转染效率为10.5%,经抗CD3抗体OKT3和IL-2共同刺激后的外周血淋巴细胞转染效率为9.9%。结论核转染方法是一种转染人外周血淋巴细胞的有效方法,转染前细胞是否受到刺激激活,对转染的效率影响不大。
Objective To determine the transfection efficiency of human peripheral blood lymphocyte with electroporation.Method The plasmid which contained EGFP gene were transfected by nucleofector transfection method,then the expression of EGFP were calculated by flow cytometry after 24 hours.Results The transfection efficiency of unstimulated lymphocytes is 8.7%.It is 10.5% for the T cells stimulated by anti-CD3/anti-CD28 antibodies and 9.9% for the T cells stimulated by anti-CD3/IL-2.Conclusion The nucleofector-based electroporation is an efficient method to transfect peripheral blood lymphocytes.The activation of T cells has no influence on transfection efficiency.
出处
《肿瘤药学》
CAS
2012年第1期28-30,共3页
Anti-Tumor Pharmacy
关键词
外周血淋巴细胞
核转染
转染效率
Peripheral blood lymphocyte
Nucleofection
Transfection efficiency
