Evaluation and optimization of DNA delivery into gliosarcoma 9L cells by a cholesterol-based cationic liposome.
This paper reports results concerning the transfection of gliosarcoma cells 9L using an original cholesterol-based cationic liposome as carrier. This cationic liposome was prepared from triethyl aminopropane carbamoyl cholesterol (TEAPC-Chol) and a helper lipid, dioleoyl phosphatidyl ethanolamine (DOPE). The used concentration of liposome was not cytotoxic as revealed by the MTT test. TEAPC-Chol/DOPE liposomes allowed the plasmids encoding reporter genes to enter the nucleus as observed both by electron microscopy and functionality tests using fluorescence detection of green fluorescent protein (GFP) and luminometric measurements of luciferase activity. By changing the cationic lipid/DNA molar charge ratio, optimal conditions were determined. Further, improvement of the transfection level has been obtained by either precondensing plasmid DNA with poly-L-lysine or by adding polyethylene glycol (PEG) in the transfection medium. The optimal conditions determined are different depending on whether the transfection is made with cells in culture or with tumors induced by subcutaneous (s.c.) injection of cells in Nude mice. For in vivo assays, a simple method to overcome the interference of haemoglobin with the chemiluminescence intensity of luciferase has been used. These results would be useful for gaining knowledge about the potential for the cationic liposome TEAPC-Chol/DOPE to transfect brain tumors efficiently.
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Laboratoire de Chimie Structurale et Spectroscopie Biomoléculaire (CSSB), CNRS FRE 2313, UFR de Médecine, Université Paris XIII, 74 rue Marcel Cachin, F93017 Bobigny Cedex, France.
Cao
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A
Briane
D
D
Lievre
N
N
Coudert
R
R
Raphael
M
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Salzmann
J l
Jl
Taillandier
E
E
D000818:Animals; D002470:Cell Survival; D002784:Cholesterol / Q000031:analogs & derivatives / Q000633:toxicity; D018014:Gene Transfer Techniques; D049452:Green Fluorescent Proteins; D008081:Liposomes; D008164:Luminescent Proteins / Q000235:genetics; D051379:Mice; D008819:Mice, Nude; D008854:Microscopy, Electron; D008858:Microscopy, Phase-Contrast; D009368:Neoplasm Transplantation; D010957:Plasmids / Q000032:analysis; D011092:Polyethylene Glycols; D011107:Polylysine; D051381:Rats; D014407:Tumor Cells, Cultured
C459406:3beta(N-(N',N',N'-triethylaminopropane)carbamoyl)cholesterol iodide; D008081:Liposomes; D008164:Luminescent Proteins; D049452:Green Fluorescent Proteins; D011107:Polylysine; D011092:Polyethylene Glycols; D002784:Cholesterol