We have been devoting our efforts to develop the useful liposomes that have high potentials of gene transfer and we aim human gene therapy for malignant glioma with cytokine genes. In our previous study, we prepared our original reverse-phase evaporation vesicles (REV) by an improved procedure of reverse-phase evaporation method. However, this procedure was very complicate. In this paper, a simple procedure for the preparation of cationic multilamellar vesicles (MLV) was introduced, and efficient expression and growth-inhibitory effect of MLV with entrapped human interferon-beta gene to glioma cells was comparable to that obtained with REV. Considering the present experiments, MLV seem to be more preferable for clinical application.