Skip to main content
SpringerLink
Log in

Electroporation as a technique for the transfer of macromolecules into mammalian cell lines

  • Published:
Journal of tissue culture methods

Summary

Numerous methods have been devised to facilitate the introduction of exogenous compounds into cells. The technique of electroporation allows the direct physical transfer of numerous kinds of molecules into essentially any cell, but the major application has been for transfection of DNA and this emphasis is recapitulated here. However, the conditions for transfer of DNA or other macromolecules are sufficiently similar that the same protocol is followed regardless. In addition, as electroporation involves a mechanism distinct from that of most other methods of transfection, it has distinct advantages and disadvantages relative to other transfection techniques. This review is designed to allow one to simplify the processes of determining whether electroporation is appropriate to a given experimental design, to indicate how to minimize the disadvantages, and to simplify the requisite process of parameter optimization required to evaluate and apply electroporation to the system of choice. Practical aspects are highlighted but the theoretical bases are discussed when relevant for application of the technique.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  1. An, G.; Hidaka, K.; Siminovitch, L. Expression of bacterialβ-galactosidase in animal cells. Mol. Cell. Biol. 2(12):1628–1632; 1982.

    Google Scholar 

  2. Anderson, M. L. M.; Spandidos, D. A.; Coggins, J. R. Electroporation of lymphoid cells: factors affecting the efficiency of transfection. J. Biochem. Biophys. Methods 22:207–222; 1991.

    Google Scholar 

  3. Andreason, G. L.; Evans, G. A. Introduction and expression of DNA molecules in eukaryotic cells by electroporation. BioTechniques 6(7):650–660; 1988.

    Google Scholar 

  4. Andreason, G. A.; Evans, G. A. Optimization of electroporation for transfection of mammalian cell lines. Anal. Biochem. 180:269–275; 1989.

    Google Scholar 

  5. Bahnson, A. B.; Boggs, S. S. Addition of serum to electroporated cells enhances survival and transfection efficiency. Biochem. Biophys. Res. Commun. 171:752–757; 1990.

    Google Scholar 

  6. Bos, R.; Nieuwenhuizen, W. Enhanced transfection of a bacterial plasmid into hybridoma cells by electroporation: application for the selection of hybrid hybridoma (quadroma) cell lines. Hybridoma 11(1):41–51; 1992.

    Google Scholar 

  7. Chang, D. C.; Gao, G.-Q.; Maxwell, B. L. High efficiency gene transfection by electroporation using a radio-frequency electric field. Biochim. Biophys. Acta 1992:153–160; 1991.

    Google Scholar 

  8. Chu, G.; Hayakawa, H.; Berg, P. Electroporation for the efficient transfection of mammalian cells with DNA. Nucleic Acids Res. 15:1311–1326; 1987.

    Google Scholar 

  9. de Wet, J. R.; Wood, K. V.; DeLuca, M., et al. Firefly luciferase gene: structure and expression in mammalian cells. Mol. Cell. Biol. 7:725–737; 1986.

    Google Scholar 

  10. Eastman, A. An improvement to the novel rapid assay for chloramphenicol acetyltransferase gene expression. BioTechniques 5:730–732; 1987.

    Google Scholar 

  11. Fraser, D.; Kiatloff-Zito, C.; Moustacchi, E. Transfection of wild-type and ‘Fanconi anemia-like’ mouse lymphoma mutant cells by electroporation. Mut. Res. 263:165–171; 1991.

    Google Scholar 

  12. Fregeau, C. J.; Bleackley, R. C. Factors influencing transient expression in cytotoxic T cells following DEAE dextran-mediated gene transfer. Somat. Cell Mol. Genet. 17(3):239–257; 1991.

    Google Scholar 

  13. Glogauer, M.; McCulloch, C. A. G. Introduction of large molecules into viable fibroblasts by electroporation: optimization of loading and identification of labeled cellular compartments. Exp. Cell Res. 200:227–234; 1992.

    Google Scholar 

  14. Goldstein, S.; Fordis, C. M.; Howard, B. H. Enhanced transfection efficiency and improved cell survival after electroporation of G2/M-synchronized cells and treatment with sodium butyrate. Nucleic Acids Res. 17:3959; 1989.

    Google Scholar 

  15. Graziadei, L.; Burfeind, P.; Bar-Sagi, D. Introduction of unlabeled proteins into living cells by electroporation and isolation of viable protein-loaded cells using dextan-fluorescein isothiocyanate as a marker for protein uptake. Anal. Biochem. 194:198–203; 1991.

    Google Scholar 

  16. Holliday, R.; Ho, T. Gene silencing in mammalian cells by uptake of 5-methyl deoxycytidine-5′-triphosphate. Somatic Cell Mol. Genet. 17(6):537–542; 1991.

    Google Scholar 

  17. Jiang, C.-K.; Connolly, D.; Blumenberg, M. Comparison of methods for transfection of human epidermal keratinocytes. J. Invest. Dermatol. 97(6):969–973; 1991.

    Google Scholar 

  18. Langlet, C.; Schmitt-Verhulst, A.-M. Electroporation of CTL clones: a useful method to investigate signalling pathways leading to the expression of effector functions. J. Immunol. Methods 151:107–115; 1992.

    Google Scholar 

  19. Neumann, E.; Schaefer-Ridder, M.; Wang, Y., et al. Gene transfer into mouse lyoma cells by electroporation in high electric fields. EMBO J. 1(7):841–845; 1982.

    Google Scholar 

  20. Nyce, J. Gene silencing in mammalian cells by direct incorporation of 5-methyl-2′-deoxycytidine 5′-triphosphate. Somatic Cell Mol. Genet. 17(6):543–550; 1991.

    Google Scholar 

  21. Okamoto, H.; Sukegawa, K.; Tomatsu, S., et al. Optimization of electroporation for transfection of human fibroblast cell lines with origin-defective SV-40 DNA: development of human transformed fibroblast cell lines with mucopolysaccharidoses (I-VII). Cell Struct. Funct. 17:123–128; 1992.

    Google Scholar 

  22. Pahl, H. L.; Burn, T. C.; Tenen, D. G. Optimization of transient transfection into human myeloid cell lines using a luciferase reporter gene. Exp. Hematol. 19:1038–1041; 1991.

    Google Scholar 

  23. Potter, H.; Weir, L.; Leder, P. Enhancer-dependent expression of human immunoglobulin genes introduced into mouse pre-B lymphocytes by electroporation. Proc. Natl. Acad. Sci. USA 81:7161–7165; 1984.

    Google Scholar 

  24. Potter, H. Electroporation in biology; methods, applications and instrumentation. Anal. Biochem. 174:361–373; 1988.

    Google Scholar 

  25. Puchalski, R. B.; Fahl, W. E. Gene transfer by electroporation, lipofection and DEAE-dextran transfection: compatibility with cell-sorting by flow cytometry. Cytometry 13:23–30; 1992.

    Google Scholar 

  26. Reid, L. H.; Shesely, E. G.; Kim, H.-S., et al. Cotransformation and gene targeting in mouse embryonic stem cells. Mol. Cell. Biol. 11(5):2769–2777; 1991.

    Google Scholar 

  27. Sambrook, J.; Fritsch, E. F.; Maniatis, T. Molecular cloning: a laboratory manual, 2nd ed. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory Press; 1989:1.33–1.46, 5.28–5.32, 16.9–16.15 and E.5:

    Google Scholar 

  28. Shigikawa, K.; Dover, W. J. Electroporation of eukaryotes and prokaryotes: a general approach to the introduction of macromolecules into cells. BioTechniques 6:742–751; 1988.

    Google Scholar 

  29. Showe, M. K.; Williams, D. L.; Showe, L. C. Quantitation of transient gene expression after electroporation. Nucleic Acids. Res. 20(12):3153–3157; 1992.

    Google Scholar 

  30. Stopper, H.; Zimmerman, U.; Neil, G. A. Increased efficiency of transfection of murine hybridoma cells with DNA by electroporation. J. Immunol Methods 109:145–151; 1988.

    Google Scholar 

  31. Takahashi, M.; Furukawa, T.; Nikkuni, K., et al. Efficient introduction of a gene into hematopoietic cells in S-phase by electroporation. Exp. Hematol. 19:343–346; 1991.

    Google Scholar 

  32. Takahashi, M.; Furukawa, T.; Saitoh, H., et al. Gene transfer into human leukemia cell lines by electroporation: experience with exponentially decaying and square wave pulse. Leuk. Res. 15(6):507–513; 1991.

    Google Scholar 

  33. Tekle, E.; Astumian, R. D.; Chock, P. B. Electroporation by using bipolar oscillating electric field: an improved method for DNA transfection of NIH 3T3 cells. Proc. Natl. Acad. Sci. USA 88:4230–2434; 1991.

    Google Scholar 

  34. Teshigawara, K.; Katsura, Y. A simple and efficient mammalian gene expression system using an EBV-based vector transfected by electroporation in G2/M phase. Nucleic Acids Res. 20(10):2607; 1992.

    Google Scholar 

  35. Toneguzzo, F.; Hayday, A. C.; Keating, A. Electric field-mediated DNA transfer: transient and stable gene expression in human and mouse hymphoid cells. Mol. Cell Biol. 6:703–706; 1986.

    Google Scholar 

  36. van den Hoff, M.; Moorman, A. F. M.; Lamers, W. H. Electroporation in ‘intracellular’ buffer increases cell survival. Nucleic Acids Res. 20(11):2902; 1992.

    Google Scholar 

  37. Zheng, Q.; Chang, D. C. High-efficiency gene transfection by in situ electroporation of cultured cells. Biochim. Biophys. Acta 1088:104–110; 1991.

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Departement de Biologie Moleculaire, Institut Pasteur, Unité de Biologie des Membranes, 25 rue du Dr. Roux, 75724, Paris cedex 15, France

    Grai L. Andreason

Authors
  1. Grai L. Andreason
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Andreason, G.L. Electroporation as a technique for the transfer of macromolecules into mammalian cell lines. Journal of Tissue Culture Methods 15, 56–62 (1993). https://doi.org/10.1007/BF01667362

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF01667362

Key words

Search

Navigation