Surfactant-mediated gene transfer for animal cells

Jun You, Masamichi Kamihira, Shinji Iijima

Research output: Contribution to journalArticlepeer-review

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
DOIs
Publication statusPublished - 1997

All Science Journal Classification (ASJC) codes

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

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