脂质体DC-Chol/DOPE对HEK293FT细胞的高效转染及其在腺病毒制备中的应用

重组病毒载体系统因为具有高效的基因转移能力得到了广泛应用,而病毒包装细胞的转染是重组病毒制备过程中的关键步骤。优化了脂质体DC-Chol/DOPE介导的转染常用的病毒包装细胞系HEK293FT的实验条件,比较了DC-Chol/DOPE、Lipofectamine2000和磷酸钙共沉淀法转染细胞的效率,并且比较了用DC-Chol/DOPE和磷酸钙共沉淀法转染293FT细胞制备重组腺病毒的结果,发现DC-Chol/DOPE对293FT细胞的转染效率以及最终收获的病毒滴度都远高于磷酸钙共沉淀法转染。所以,利用DC-Chol/DOPE转染293FT细胞制备重组病毒是一种简单、高效、成本低廉的方法。

Production of Adenoviral Vector Using Liposome DC-Chol/DOPE Transfection of HEK293FT Cells

Ability of viral vectors to achieve efficient gene transfer in most of mammalian cell types makes them useful tools in many fields. The transfection of virus packaging cells is a most critical step for high-titer viral vector production. In order to transfer gene efficiently to commonly used packaging cell line HEK293FT, transfection parameters for cationic liposome DC-Chol/DOPE were optimized, including DNA amount, liposome-to-DNA weight ratio and transfection buffer. The efficiency of DNA transfer into 293FT cells were compared using DC-Chol/DOPE liposome, Lipofectamine2000 and calcium phosphate. The transfection efficiency of DC-Chol/DOPE is higer than calcium phosphate, and is almost equal to Lipofectamine2000. The adenovirus yield by DC-Chol/DOPE and calcium phosphate transfection were compared also. The virus yield from DC-Chol/DOPE trasfection was much more than calcium phosphate. Therefore, DC-Chol/DOPE is a high efficient, easy to use and inexpensive transfection reagent which is suitable for production of viral vectors.