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原代树突细胞基因递送系统的构建与应用

于泽萱, 鞠曹云, 张灿

于泽萱, 鞠曹云, 张灿. 原代树突细胞基因递送系统的构建与应用[J]. 中国药科大学学报, 2021, 52(4): 438-446. DOI: 10.11665/j.issn.1000-5048.20210406
引用本文: 于泽萱, 鞠曹云, 张灿. 原代树突细胞基因递送系统的构建与应用[J]. 中国药科大学学报, 2021, 52(4): 438-446. DOI: 10.11665/j.issn.1000-5048.20210406
YU Zexuan, JU Caoyun, ZHANG Can. Construction and application of gene delivery systems for primary dendritic cells[J]. Journal of China Pharmaceutical University, 2021, 52(4): 438-446. DOI: 10.11665/j.issn.1000-5048.20210406
Citation: YU Zexuan, JU Caoyun, ZHANG Can. Construction and application of gene delivery systems for primary dendritic cells[J]. Journal of China Pharmaceutical University, 2021, 52(4): 438-446. DOI: 10.11665/j.issn.1000-5048.20210406

原代树突细胞基因递送系统的构建与应用

基金项目: 国家自然科学基金资助项目(No.82073785,No.81930099,No.81773664)

Construction and application of gene delivery systems for primary dendritic cells

Funds: This study was supported by the National Natural Science Foundation of China (No. 82073785, No.81930099, No.81773664)
  • 摘要: 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。
    Abstract: 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.
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出版历程
  • 收稿日期:  2021-04-24
  • 修回日期:  2021-06-28
  • 刊出日期:  2021-08-24

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