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目的考察非病毒载体右旋糖酐-精胺在多种细胞上的转染效果,构建新型基因传递体系。方法制备右旋糖酐-精胺/DNA复合物,在体外考察粒径、Zeta电位等指标。运用常规转染方法在多种肿瘤细胞(HepG2、HeLa、A549)以及骨髓间充质干细胞(MSCs)上进行转染实验,考察血清存在与否对转染效率的影响。合成阴离子化明胶,构建反向转染体系,比较常规转染方法与反向转染方法下右旋糖酐-精胺转染效果的差异。结果右旋糖酐-精胺可以有效转染细胞,相比正向转染,反向转染体系中血清存在对基因转染效率的影响较小。结论右旋糖酐-精胺是一种有潜在应用价值的非病毒基因载体,反向转染体系是一种有前途的基因传递方法。
Objective To investigate the transfection efficiency of non-viral vector dextran-spermine on various cells and to construct a novel gene delivery system. Methods Dextran - spermine / DNA complex was prepared and its particle size and Zeta potential were investigated in vitro. Transfection experiments were performed on a variety of tumor cells (HepG2, HeLa, A549) and bone marrow mesenchymal stem cells (MSCs) using conventional transfection methods to investigate the effect of serum on the transfection efficiency. Anionic gelatin was synthesized and the reverse transfection system was constructed. The difference of dextran-spermine transfection efficiency between conventional transfection method and reverse transfection method was compared. Results Dextran-spermine could effectively transfect cells. Compared with the positive transfection, the presence of serum in the reverse transfection system had little effect on gene transfection efficiency. Conclusion Dextran-spermine is a potential non-viral gene vector, reverse transfection system is a promising method of gene delivery.