Liposomal mediated transfection of mature rat astrocytes in vitro: a comparison of ten reagents.
Ten cationic liposomes were tested for their ability to transfect mature, astrocyte monolayers in vitro using the eucaryotic expression vector plasmid p cytomegalovirus (CMV)-beta. Liposomal agents were examined for optimum length of exposure and optimum cDNA/lipid ratios. Lipofectin demonstrated the highest transfection efficiencies of all agents tested (3.3%). When examined at 3 days following transfection, 24-h exposures yielded higher efficiencies compared to 6 h exposures (1.9%, P=0.07). Although expression appeared to decline by up to 80%, positive cells were still detected up to 2 weeks after transfection with all reagents. Lipofectin represents a useful tool for transfecting mature astrocytes for investigation of gene transfer in vitro.
McKalip
D
D
UCSF Department of Neurological Surgery, San Francisco, CA 94066, USA. mckalip@neurosurg.ucsf.edu
Grenier
M
M
Choy
M
M
D000818:Animals; D001253:Astrocytes / Q000502:physiology; D002478:Cells, Cultured; D003587:Cytomegalovirus / Q000235:genetics; D005229:Fatty Acids, Monounsaturated / Q000494:pharmacology*; D005456:Fluorescent Dyes / Q000494:pharmacology*; D008081:Liposomes; D010714:Phosphatidylethanolamines / Q000494:pharmacology*; D000644:Quaternary Ammonium Compounds / Q000494:pharmacology*; D051381:Rats; D017207:Rats, Sprague-Dawley; D014162:Transfection / Q000379:methods*
D005229:Fatty Acids, Monounsaturated; D005456:Fluorescent Dyes; D008081:Liposomes; D010714:Phosphatidylethanolamines; D000644:Quaternary Ammonium Compounds; C020888:1,2-dielaidoylphosphatidylethanolamine; C070046:1,2-dioleoyloxy-3-(trimethylammonium)propane