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Transfection of insect cell lines using polyethylenimine

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Abstract

Insect cell lines have been widely used in recombinant baculovirus expression systems and transient gene expression studies. Critical to these applications have been the transfection of foreign DNA. This has been frequently done using labor intensive and cytotoxic liposome-based transfection reagents. In the current study we have optimized a new kind of polyethylenimine-based DNA transfection reagent on the Spodoptera frugiperda Sf9 insect cell line. A plasmid vector that transiently expresses green fluorescent protein (GFP) was effectively delivered into Sf9 cells. A transfection efficiency of 54% and cell viability of 85–90% were obtained for Sf9 cells. The developed transfection protocol has now been successfully used to transfect eight insect cell lines derived from Bombyx mori, Trichoplusia ni, Helicoverpa zea, Heliothis virescens and S. frugiperda with GFP and GUS with transfection efficiencies of at least 45%. This method provides high heterologous protein expression levels, transfection efficacy and cell viability, and could be used for transient gene expression in other lepidopteran cell lines.

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  1. GS Laboratory Equipment, 275 Aiken Rd., Asheville, NC 28804, USA.

  2. Faster, 46 Via Vespucci, Ferrara 44100, Italy.

  3. Grant Instruments (Cambridge) Limited, 29 Station Rd., Shepreth, Royston, Herts SG8 6PZ, England.

  4. Leica Microsystems Wetzlar GmbH, 17–37 Ernst-Leitz Str., Wetzlar 35578, Germany.

  5. Rainin Instrument Co., Inc., P.O. Box 4026, Mack Rd., Woburn, MA 01888–4026, USA.

  6. Drummond Scientific Co., 500 Parkway, P.O. Box 700, Broomall, PA 19008, USA.

  7. Stuart Scientific Co., Staffordshire, UK.

  8. Eppendorf AG, 1 Barkhausenweg, Hamburg 22339, Germany.

  9. Hausser Scientific Co., 935 Horsham Rd., Suite C, Horsham, PA 19044–1286, USA.

  10. Leica Microsystems Imaging Solution Ltd. PO Box 86, 515 Coldhams Lane, Cambridge CB1 3XJ, UK.

  11. Amersham Biosciences Corp., P.O. Box 1327, 800 Centennial Ave., Piscataway, NJ 08855–1327, USA.

  12. Adobe Systems Incorporated, San Jose, CA 95110, USA.

  13. Microsoft Corp., One Microsoft way, Redmond, WA 9052–6399, USA.

  14. Life Technologies, 3175 Staley Rd., Grand Island, NY 14072, USA.

  15. Sigma, P.O. Box 14508, St. Louis, MO63178, USA.

  16. Fermentas Inc., 7520 Conneley Drive, Suite A, Hanover, MD 21076, USA.

  17. Greiner, GmBH D-72636 Frickenhausen, West Germany.

  18. Kimble/Kontes, Vineland, NJ, USA.

  19. Falcon brand, Beckon Dickenson and Co, Lincoln Park, NJ 07035, USA.

  20. Costar, 205 Broadway, Cambridge, MA 02139, USA.

  21. ISC BioExpress, 420 North Kays Dr., Kaysville, UT 84037, USA.

  22. Fisher Scientific, 3970 Johns Creek Court Suite 500, Suwanee, GA 30024, USA.

  23. Pall Gelman Laboratory, 600 South Wagner Rd., Ann Arbor, MI 48103–9019 USA.

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Acknowledgements

Mention of trade names or commercial products in this publication is solely for the purpose of providing specific information and does not imply recommendation or endorsement by the US Department of Agriculture. A Science & Technology Centre of Ukraine grant (STCU project 119) supported this project and was part of collaboration between the Uzbekistan Academy of Sciences of Uzbekistan, UC Davis and the USDA.

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Author notes

  1. I. D. Ogay & O. A. Lihoradova

    Present address: Institute of the Chemistry of Plant Substances, Uzbek Academy of Science, Tashkent, 700170, Uzbekistan

  2. J. M. Slack

    Present address: Great Lakes Forestry Centre, Canadian Forestry Service, Natural Resources Canada, 1219 Queen St, P6A 2E5, Sault Ste Marie, ON, Canada

Authors and Affiliations

  1. Institute of Genetics and Plant Experimental Biology, Uzbek Academy of Science Tashkent region, Qibray district, Yuqori-Yuz, 702151, Uzbekistan

    I. D. Ogay, O. A. Lihoradova & A. A. Abdukarimov

  2. Institute of the Chemistry of Plant Substances, Uzbek Academy of Science, Tashkent, 700170, Uzbekistan

    Sh. S. Azimova

  3. Insect Biocontrol Laboratory, USDA/ARS, BARC-West, Bldg 011A, Rm 214, Beltsville, MD, 20852-2350, USA

    J. M. Slack & D. E. Lynn

Authors
  1. I. D. Ogay
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  2. O. A. Lihoradova
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  3. Sh. S. Azimova
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  4. A. A. Abdukarimov
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  5. J. M. Slack
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  6. D. E. Lynn
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Correspondence to D. E. Lynn.

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As a U.S. Government employee, the article is not copyrightable.

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Ogay, I.D., Lihoradova, O.A., Azimova, S.S. et al. Transfection of insect cell lines using polyethylenimine. Cytotechnology 51, 89–98 (2006). https://doi.org/10.1007/s10616-006-9022-7

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  • DOI: https://doi.org/10.1007/s10616-006-9022-7

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