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 evaluate the transfection effect of dextran-spermine and to develop a novel gene delivery system.METHODS Dextran-spermine was complexed with the plasmid DNA coding for luciferase,then the particle size and Zeta potentials of the nanoparticles were investigated.Dextran-spermine /pDNA complexes were used to transfect tumor cells and mesenchymal stem cells either in the presence or absence of serum.Anionized gelatin was synthesized and used to construct reverse transfection system.Gene expression with reverse transfection method was compared with the conventional transfection method.RESULTS Dextran-spermine /pDNA complexes can effectively transfect cells.In contrast to the conventional transfection method,the reverse transfection method permitted the transfection of cells in the presence of serum.CONCLUSION Dextran-spermine is a promising gene carrier.The reverse transfection system has reduced the negative influence of serum,and will be a promising gene delivery method.