One of the major challenges for successful gene therapy is improving the transfection efficiency of non-viral vectors. Magnetic nanoparticles (MNPs) have been developed as enhancers of non-viral vehicles. We prepared MNPs and modified them with polyethyleneimine (PEI), citric acid (CA) or carboxylmethyl-dextran (CMD). Both positively charged MNPs (MNPs@PEI) and negatively charged MNPs (MNPs@CA, MNPs@CMD) could spontaneously form transfection complexes (magnetofectins) with plasmid DNA and PEI/liposome via electrostatic self-assembly. Our results showed as-prepared magnetofectins apparently enhanced PEI/liposome transfection efficiency and/or gene expression level into COS-7 cells with reduced transfection time from 4 h to 15 min under a magnetic field in vitro. Meanwhile, the effect of magnetofection was cell line-dependant. These results suggest that charged MNPs could improve transfection efficiency for non-viral vectors by simply mixing with them and by exerting a magnetic force. Thus such MNPs provide a convenient platform for further applications of gene delivery. In order to combine MNPs with non-viral vectors, MNPs were coated with PEI [4,5] or cationic liposomes [6]. In fact, according to the mechanism study on magnetofection, MNPs play the role of driving vector/pDNA complexes to cell surface in magnetofection and do not directly affect the endocytic uptake mechanism [4]. Magnetofection requires MNPs to have sufficient electric potentials at the surface, so it is not necessary to modify MNPs with these gene vectors according to our previous study [7]. Our experimental evidences indicate that MNPs could not enter nucleus once they were internalized by the cells. MNPs would probably have to be separated from vector/pDNA complexes after entering cytoplasm, although detailed mechanism requires further investigation. In this study, we aim to further demonstrate charged MNPs, both positively charged and negatively charged MNPs, can improve transfection efficiency for non-viral vectors by simply mixing with them via electrostatic self-assembly.In this paper, we introduced the preparation, cytotoxicity of magnetofectins as prepared via electrostatic self-assembly, and evaluated the performance of these magnetofectins in magnetofection as follows: (1) to determine if MNPs@PEI can improve PEI reagent transfection; (2) to determine if negatively charged MNPs@CA and MNPs@CMD can improve PEI reagent transfection; (3) to determine if charged