Experimental design optimization of filamentous phage transfection into mammalian cells by cationic lipids.
A previous study showed that filamentous phage could be efficiently transfected into mammalian cells in the presence of the cationic lipid Transfectam. In the present study, we used an experimental plan based on a uniform network (Doehlert) matrix to estimate optimal transfection conditions in two different cell lines, CHO and Cos-7. Using the cationic lipid RPR120535b as a model, we show that optimal conditions can be determined much more readily than with standard response curves. Under optimal conditions as analyzed by FACS, up to 60% of Cos-7 and 50% of CHO cells can be transfected. Furthermore, a comparison of different lipids (Transfectam, RPR120535b, TC1-12 and GAP-DLRIE/DOPE) suggests that lipids with multiple amine groups are more efficient for the transfection of filamentous phage.
Aujame
L
L
luc.aujame@aventis.com
Seguin
D
D
Droy
C
C
Hessler
C
C
D000818:Animals; D001435:Bacteriophages / Q000235:genetics*; D016466:CHO Cells; D019556:COS Cells; D006224:Cricetinae; D008055:Lipids / Q000494:pharmacology*; D012107:Research Design; D014162:Transfection
D008055:Lipids