Surfactant-mediated gene transfer for animal cells

Jun You, Masamichi Kamihira, Shinji Iijima

Research output: Contribution to journalArticle

17 Citations (Scopus)

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.

Original languageEnglish
Pages (from-to)45-52
Number of pages8
JournalCytotechnology
Volume25
Issue number1-3
Publication statusPublished - Dec 1 1997
Externally publishedYes

Fingerprint

Gene transfer
Surface-Active Agents
Lipids
Animals
Surface active agents
Cells
Transfection
Genes
DNA
Cationic surfactants
Galactosidases
Polysorbates
Nonionic surfactants
Cytotoxicity
Plasmids
Weights and Measures
Temperature
Water

All Science Journal Classification (ASJC) codes

  • Biotechnology
  • Bioengineering
  • Biomedical Engineering
  • Clinical Biochemistry
  • Cell Biology

Cite this

Surfactant-mediated gene transfer for animal cells. / You, Jun; Kamihira, Masamichi; Iijima, Shinji.

In: Cytotechnology, Vol. 25, No. 1-3, 01.12.1997, p. 45-52.

Research output: Contribution to journalArticle

You, J, Kamihira, M & Iijima, S 1997, 'Surfactant-mediated gene transfer for animal cells', Cytotechnology, vol. 25, no. 1-3, pp. 45-52.
You, Jun ; Kamihira, Masamichi ; Iijima, Shinji. / Surfactant-mediated gene transfer for animal cells. In: Cytotechnology. 1997 ; Vol. 25, No. 1-3. pp. 45-52.
@article{ea2d02e82bc7421bb208f200c3d9e542,
title = "Surfactant-mediated gene transfer for animal cells",
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.",
author = "Jun You and Masamichi Kamihira and Shinji Iijima",
year = "1997",
month = "12",
day = "1",
language = "English",
volume = "25",
pages = "45--52",
journal = "Cytotechnology",
issn = "0920-9069",
publisher = "Springer Netherlands",
number = "1-3",

}

TY - JOUR

T1 - Surfactant-mediated gene transfer for animal cells

AU - You, Jun

AU - Kamihira, Masamichi

AU - Iijima, Shinji

PY - 1997/12/1

Y1 - 1997/12/1

N2 - 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.

AB - 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.

UR - http://www.scopus.com/inward/record.url?scp=0031416195&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0031416195&partnerID=8YFLogxK

M3 - Article

C2 - 22358878

AN - SCOPUS:0031416195

VL - 25

SP - 45

EP - 52

JO - Cytotechnology

JF - Cytotechnology

SN - 0920-9069

IS - 1-3

ER -