Although the cell viability after treatment with pDNA/AFliposomes and pDNA/CyDs/AF-liposomes for 3 h slightly decreased as a charge ratio of AF-liposomes/pDNA was increased in both HepG2 and NIH3T3 cells, that is, more than 80% of cell viability after application of pDNA/CyDs/AF-liposomes was observed at the charge ratio of 1.6 used in the transfection study as described above. These results suggest that pDNA/CyDs/AF-liposomes have great advantages as a nonviral vector, that is, superior transfection efficiency and less cytotoxicity, and the enhancing effect of γ-CyD on gene transfer activity of pDNA/AFliposomes is not associated with cytotoxicity.