Abstract
A commercially available cationic surfactant, dimethyl-dioctadecyl ammonium bromide (DDAB), was used for making lipid vesicles. DDAB easily dissolved in water at 60 °C and formed lipid vesicles at room temperature. The lipid vesicles showed very low cytotoxicity compared with other cationic surfactants. After the lipid vesicles were mixed with plasmid DNA solution, the solution was added to mammalian cells. The addition of a nonionic surfactant (Tween 80) to the cationic lipid vesicles at the weight ratio of 1:1 enhanced transfection efficiency. Adding more or less than the optimal amounts of DNA and lipid vesicles resulted in decreased transfection efficiency. With the optimal amounts of DNA (pCMVβ) and lipid vesicles, about 90–95% of CHO-K1 and BHK-21C13 cells transiently expressed β-galactosidase activity 24 h after transfection. By this procedure, stable transformants around 105 cells corresponding to 10% efficiency could be obtained by one batch transfection.
Similar content being viewed by others
References
Bennett MJ, Nantz MH, Balasubramaniam RP, Gruenert DC and Malone RW (1995) Cholesterol enhances cationic liposome-mediated DNA transfection of human respiratory epithelial cells. Biosci Rep 15: 47–53.
Bond VC and Wold B (1987) Poly-L-ornithine-mediated transformation of mammalian cells. Mol Cell Biol 7: 2286–2293.
Felgner PL, Gadek TR, Holm M, Roman R, Chan HW, Wenz M, Northrop JP, Ringold GM and Danielsen M (1987) Lipofection: a highly efficient, lipid-mediated DNA-transfection procedure. Proc Natl Acad Sci USA 84: 7413–7417.
Felgner PL and Ringold GM (1989) Cationic liposome-mediated transfection. Nature 337: 387–388.
Hall CV, Jacob PE, Ringold GM and Lee F (1983) Expression and regulation of Escherichia coli lacZ gene fusions in mammalian cells. J Mol Appl Genet 2: 101–109.
Holmen SL, Vanbrockin MW, Eversole RR, Stapleton SR and Ginsberg LC (1995) Efficient lipid-mediated transfection of DNA into primary rat hepatocytes. In Vitro Cell Dev Biol 31: 347–351.
Klebe RJ, Harriss JV, Hanson DP and Gauntt CJ (1984) High-efficiency polyethylene glycol-mediated transformation of mammalian cells. Som Cell Mol Genet 10: 495–502.
Lappalainen K, Jaaskelainen I, Syrjanen K, Urtti A and Syrjanen S (1994) Comparison of cell proliferation and toxicity assays using two cationic liposomes. Pharmaceu Res 11: 1127–1131.
LeVentis R and Silvius JR (1990) Interactions of mammalian cells with lipid dispersions containing novel metabolizable cationic amphiles. Biochim Biophys Acta 1023: 124–132.
Macgregor GR and Caskey CT (1989) Construction of plasmids that express E. coli β-galactosidase in mammalian cells. Nucleic Acids Res 17: 2365.
Malone RW, Felgner PL and Verma IM (1989) Cationic liposome-mediated RNA transfection. Proc Natl Acad Sci USA 86: 6077–6081.
Mannino RJ and Fould-Fogerite S (1988) Liposome mediated gene transfer. Biotechniques 6: 682–690.
Nabel GJ, Nabel EG, Yang ZY, Fox B, Plautz GE, Gao X, Huang L, Shu S, Gordon D and Chang AE (1993) Direct gene transfer with DNA-liposome complexes in melanoma: Expression, biologic activity, and lack of toxicity in humans. Proc Natl Acad Sci USA 1023: 124–132.
Potter H, Weir L and Leder P (1984) Enhancer-dependent expression of human kappa immunoglobulin genes introduced into mouse pre-B lymphocytes by electroporation. Proc Natl Acad Sci USA 81: 7161–7165.
Rose JK, Buonocore L and Whitt MA (1991) A new cationic liposome reagent mediating nearly quantitative transfection of animal cells. Biotechniques 10: 520–525.
Yagi K, Noda H, Kurono M and Ohishi N (1993) Efficient gene transfer with less cytotoxicity by means of cationic multilamellar liposomes. Biochem Biophys Res Commun 196: 1042–1048.
Rights and permissions
About this article
Cite this article
You, J., Kamihira, M. & Iijima, S. Surfactant-mediated gene transfer for animal cells. Cytotechnology 25, 45–52 (1997). https://doi.org/10.1023/A:1007955631313
Issue Date:
DOI: https://doi.org/10.1023/A:1007955631313