PURPOSE: To reduce damage to cells in transducing a gene and enable obtaining of tranformant cells in a large amount by preparing a multilamellar liposome by vortex treatment.
CONSTITUTION: N-(α-Trimethylammonioacetyl)-didodecyl-D-glutamate chloride, dilauroylphosphatidylcholine and dioleoylphosphatidyldiethanolamine are weighed so as to provide 1:2:2 molar ratio and dissolved in a solvent. The solvent is then removed by using a rotary evaporator to form a lipid thin film. A solution of a genetic DNA is then added to carry out vortex treatment. Thereby, a multilamellar liposome [multilamellar vesicle(MLV)] capturing the gene is formed. In order to efficiently perform genetic transduction into cells, it is necessary to bring the MLV into sufficient contact with the cells. The optimum contact time therefor varies with the kind of the cells into which the gene is transduced. Thereby, preexperiments are preferably carried out before the transduction of the gene to preset the time without exhibiting cytotoxicity.
NODA HITOSHI
OISHI SHIGEKO
KURONO MASATSUNE
