原代树突细胞基因递送系统的构建与应用

目前,安全有效的基因递送系统应用于原代树突细胞（dendritic cell,DC）转染尚未见报道,本研究针对原代DC构建基于脂质体的基因递送系统,优化了制备方法以提高原代DC的转染效率。通过共孵育法、乙醇注入法、鱼精蛋白复合法分别制备含不同阳离子脂质的脂质体/siRNA复合物,并以粒径、电位、对基因药物siRNA的包载能力、安全性、稳定性、DC的摄取效率和基因沉默效率为考察指标,评价并筛选出具有高转染效率的基因递送系统。与市售制剂Lipo2000相比,利用共孵育法制备的赖氨酸谷氨酸双油醇酯（OA2）递送系统的DC摄取效率提高约35%,基因沉默效率提高约10倍,且细胞存活率比Lipo2000提高20%,具有良好的DC基因转染效率和体外安全性。本研究为原代DC提供了一种安全有效、制备简单的基因递送载体平台,具有良好的应用前景。

Construction and application of gene delivery systems for primary dendritic cells

Nowadays, there is still no mature gene delivery system for safe and effective transfection on primary dendritic cells (DC). Herein, we constructed a liposome-based gene delivery system for primary DCs and optimized the preparation method to improve the transfection efficiency of siRNA on primary DCs. In this study, different methods, including co-incubation method, ethanol injection method, and protamine compound method, were used to prepare liposome/siRNA complexes based on different cationic lipids. Moreover, particle size, zeta potential, siRNA loading capacity, safety, stability, uptake efficiency and gene silencing efficiency of various liposome/siRNA complexes were detected to screen the optimal cationic lipid as well as its preparation method. We demonstrated that the OA2/siRNA delivery system prepared by the co-incubation method exhibited the best safety, uptake efficiency and gene silencing effect, compared to other siRNA delivery systems including the commercial Lipo2000. In summary, we provide a safe and effective gene delivery vector for primary DC cells through simple preparation method, which could also offer a gene delivery platform for other immune cells.