Polyplexes are polyelectrolyte complexes of DNA and polycations, designed for potential gene delivery. We investigated the properties of new polyplexes formed from cholesterol-modified polycations and DNA. Three complexes were tested; their cholesterol contents were 1.4, 6.3, and 8.7 mol %. UV spectroscopy and fluorescence assay using ethidium bromide proved the formation of polyplexes. The kinetics of turbidity of polyplexes solutions in physiological solution showed that the colloid stability of polyplexes increases with increasing content of cholesterol in polycations. Dynamic, static, and electrophoretic light scattering, small-angle X-ray scattering, and atomic force microscopy were used for characterization of polyplexes. The observed hydrodynamic radii of polyplexes were in the range of 30-60 nm; they were related to the polycation/DNA ratio and hydrophobicity of the used polycations (the cholesterol content). The properties of polyplex particles depend, in addition to polycation structure, on the rate of polycation addition to DNA solutions.