The analysis of serum effects on structure, size and toxicity of DDAB-DOPE and DC-Chol-DOPE lipoplexes contributes to explain their different transfection efficiency.
The effect of serum on structural properties of dimethyl-dioctadecyl-ammonium bromide (DDAB)-1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE) liposomes and DDAB-DOPE/DNA lipoplexes has been investigated by energy dispersive X-ray diffraction (EDXD) technique, at different cationic lipid/DNA weight ratios (rho). The role of serum on the size of lipoplexes has also been studied by dynamic light scattering. Lipoplex transfection efficiency (TE) as a function of rho, and lipoplex toxicity to C6 rat glioma cells have been evaluated in Dulbecco's Modified Eagle Medium (DMEM) with and without serum. A multi-parametric analysis concerning the role of size, structure and cytotoxicity on transfection efficiency contributes to explain the experimental observation that 3beta-[N-(N',N'-dimethylaminoethane)carbamoyl]-cholesterol (DC-Chol)-DOPE/DNA transfect C6 cells better than DDAB-DOPE/DNA lipoplexes.
Esposito
Claudio
C
Istituto di Neurobiologia e Medicina Molecolare-CNR, Via Fosso del Cavaliere 100, 00133 Roma, Italy.
Generosi
Johanna
J
Mossa
Giuseppe
G
Masotti
Andrea
A
Castellano
Agostina Congiu
AC
D000818:Animals; D002412:Cations; D045744:Cell Line, Tumor / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D002784:Cholesterol / Q000031:analogs & derivatives* / Q000737:chemistry / Q000633:toxicity; D004273:DNA, Neoplasm / Q000737:chemistry* / Q000235:genetics; D018014:Gene Transfer Techniques; D005910:Glioma / Q000188:drug therapy; D008055:Lipids / Q000737:chemistry*; D008081:Liposomes / Q000737:chemistry*; D010714:Phosphatidylethanolamines / Q000737:chemistry*; D000644:Quaternary Ammonium Compounds / Q000737:chemistry / Q000633:toxicity*; D051381:Rats; D044967:Serum / Q000737:chemistry*; D013329:Structure-Activity Relationship; D018675:Toxicity Tests; D014162:Transfection; D014867:Water / Q000737:chemistry; D014961:X-Ray Diffraction
C410262:1,2-dioleoyl-glycero-3-phosphatidyl ethanolamine; D002412:Cations; D004273:DNA, Neoplasm; D008055:Lipids; D008081:Liposomes; D010714:Phosphatidylethanolamines; D000644:Quaternary Ammonium Compounds; D014867:Water; C046112:didodecyldimethylammonium; C070648:3-(N-(N',N'-dimethylaminoethane)carbamoyl)cholesterol; D002784:Cholesterol