OPTIMIZING LIPOSOME-MEDIATED GENE-TRANSFER IN PRIMARY RAT SEPTOHIPPOCAMPAL CELL-CULTURES

Although liposomes have been widely employed to transfect DNA into a v ariety of cell types, no previous studies have systematically examined conditions producing optimal liposomal-mediated transfection of DNA i nto central nervous system (CNS) cells. Thus, we used the beta-galacto sidase (beta-gal) reporter gene to examine factors influencing the eff iciency of liposome-mediated gene transfection in CNS cell cultures. O ur results indicate that without increasing the amounts of DNA, increa sed liposome concentrations within certain limits enhanced transfectio n efficiency. However, higher liposome levels could produce cell lysis , Without increasing liposome concentrations, increased amounts of DNA did not improve transfection efficiency. Employing the optimal concen tration (1 mu g DNA/3 mu l liposomes/well), beta-gal gene expression w as sustained for at least two weeks after transfection in primary sept o-hippocampal cultures.