Cationic multilamellar liposome-mediated gene transfer into primary myoblasts.
To transfer foreign genes into myoblasts in primary culture, we found cationic multilamellar liposomes to be a useful mediator. When the cells were transfected with 2 micrograms of the plasmid pRSV-luc encapsulated into 50 nmol of our cationic multilamellar liposomes that had been prepared from N-(alpha-trimethylammonioacetyl)-didodecyl-D-glutamate chloride, dilauroyl phosphatidylcholine, and dioleoyl phosphatidylethanolamine in a molar ratio of 1:2:2, luciferase was expressed with high efficiency without cytotoxicity. When the cells were transfected with the plasmid pRSV-lacZ encapsulated into the same liposomes, 0.7% of the cultured myoblasts expressed beta-galactosidase without cytotoxicity. This is the first successful instance of introducing foreign genes into primary cultures of myoblasts by means of liposomes.
Kojima
H
H
Institute of Applied Biochemistry, Yagi Memorial Park, Gifu, Japan.
Ohishi
N
N
Takamori
M
M
Yagi
K
K
D000818:Animals; D000831:Animals, Newborn; D002478:Cells, Cultured; D004337:Drug Carriers; D018014:Gene Transfer Techniques; D005971:Glutamates; D008081:Liposomes; D008156:Luciferases / Q000032:analysis / Q000096:biosynthesis; D018482:Muscle, Skeletal / Q000378:metabolism*; D010713:Phosphatidylcholines; D010714:Phosphatidylethanolamines; D010957:Plasmids; D051381:Rats; D011994:Recombinant Proteins / Q000032:analysis / Q000096:biosynthesis; D014162:Transfection / Q000379:methods*
D004337:Drug Carriers; D005971:Glutamates; D008081:Liposomes; D010713:Phosphatidylcholines; D010714:Phosphatidylethanolamines; D011994:Recombinant Proteins; C084512:didodecyl N-(alpha-trimethylammonioacetyl)glutamate; C015153:1,2-dilauroylphosphatidylcholine; C020888:1,2-dielaidoylphosphatidylethanolamine; D008156:Luciferases