A new cationic liposome for efficient gene delivery with serum into cultured human cells: a quantitative analysis using two independent fluorescent probes

Cationic liposomes are useful to transfer genes into eukaryotic cells in vi tro and in vivo. However, liposomes with good transfection efficiency are o ften cytotoxic, and also require serum-free conditions for optimal activity . In this report, we describe a new formulation of cationic liposome contai ning DC-6-14, O,O'-ditetradecanoyl-N-(alpha-trimethylammonioacethyl)diethan olamine chloride, dioleoylphosphatidylethanolamine and cholesterol for gene delivery into cultured human cells. This liposome, dispersed in 5% serum-c ontaining growth medium, efficiently delivered a plasmid DNA for GFP (green fluorescent protein) into more than 80% of the cultured human cell hybrids derived from HeLa cells and normal fibroblasts. Flow cytometric analysis r evealed that the efficiency of the GFP gene expression was 40-50% in a tumo r-suppressed cell hybrid, while it was greatly reduced in the tumorigenic c ounterpart. The enhanced GFP expression in tumor-suppressed cell hybrids wa s quantitatively well correlated with a prolonged presence of the plasmid D NA, which had been labeled with another fluorescent probe, ethidium monoazi de, within the cells, These results suggest that a newly developed cationic liposome is useful for gene delivery in serum-containing medium into human cells and the stability of the plasmid DNA inside the cell is a crucial st ep in this liposome-mediated gene expression. The mechanisms by which catio nic liposome mediates gene transfer into eukaryotic cells are also discusse d. (C) 2000 Elsevier Science B.V. All rights reserved.