Analysis and Comparison of Electrical Pulse Parameters for Gene Electrotransfer of Two Different Cell Lines
Knowledge of the parameters which influence the efficiency of gene electrotransfer has importance for practical implementation of electrotransfection for gene therapy as well as for better understanding of the underlying mechanism. The focus of this study was to analyze the differences in gene electrotransfer and membrane electropermeabilization between plated cells and cells in a suspension in two different cell lines (CHO and B16F1). Furthermore, we determined the viability and critical induced transmembrane voltage (ITVc) for both cell lines. In plated cells we obtained relatively little difference in electropermeabilization and gene electrotransfection between CHO and B16F1 cells. However, significant differences between the two cell lines were observed in a suspension. CHO cells exhibited a much higher gene electrotransfection rate compared to B16F1 cells, whereas B16F1 cells reached maximum electropermeabilization at lower electric fields than CHO cells. Both in a suspension and on plated cells, CHO cells had a slightly better survival rate at higher electric fields than B16F1 cells. Calculation of ITVc in a suspension showed that, for both electropermeabilization and gene electrotransfection, CHO cells have lower ITVc than B16F1 cells. In all cases, ITVc for electropermeabilization was lower than ITVc for gene electrotransfer, which is in agreement with other studies. Our results show that there is a marked difference in the efficiency of gene electrotransfer between suspended and plated cells.
KeywordsGene electrotransfer Electropermeabilization Induced transmembrane voltage ITVc CHO B16F1 Suspension Plated cell
This research was supported by the Slovenian Research Agency under grants J2-9770 and P2-0249. We thank also Rosana Hudej (Faculty of Chemistry and Chemical Technology, University of Ljubljana) and Marko Ušaj (Faculty of Electrical Engineering, University of Ljubljana) for help in experimental procedures.
- Corovic S, Zupanic A, Miklavcic D (2008) Numerical modeling and optimization of electric field distribution in subcutaneous tumor treated with electrochemotherapy using needle electrodes., 36:1665–1672Google Scholar
- Hacein-Bey-Abina S, Von Kalle C, Schmidt M, McCormack MP, Wulffraat N, Leboulch P, Lim A, Osborne CS, Pawliuk R, Morillon E, Sorensen R, Forster A, Fraser P, Cohen JI, de Saint Basile G, Alexander I, Wintergerst U, Frebourg T, Aurias A, Stoppa-Lyonnet D, Romana S, Radford-Weiss I, Gross F, Valensi F, Delabesse E, Macintyre E, Sigaux F, Soulier J, Leiva LE, Wissler M, Prinz C, Rabbitts TH, Le Deist F, Fischer A, Cavazzana-Calvo M (2003) LMO2-associated clonal T cell proliferation in two patients after gene therapy for SCID-X1. Science 302:415–419CrossRefPubMedGoogle Scholar
- Mitsuyasu RT, Merigan TC, Carr A, Zack JA, Winters MA, Workman C, Bloch M, Lalezari J, Becker S, Thornton L, Akil B, Khanlou H, Finlayson R, McFarlane R, Smith DE, Garsia R, Ma D, Law M, Murray JM, von Kalle C, Ely JA, Patino SM, Knop AE, Wong P, Todd AV, Haughton M, Fuery C, Macpherson JL, Symonds GP, Evans LA, Pond SM, Cooper DA (2009) Phase 2 gene therapy trial of an anti-HIV ribozyme in autologous CD34+ cells. Nat Med 15:285–292CrossRefPubMedGoogle Scholar
- Neumann E, Schaefer-Ridder M, Wang Y, Hofschneider PH (1982) Gene transfer into mouse lymoma cells by electroporation in high electric fields. EMBO J 7:841–845Google Scholar
- Neumann E, Sowers AE, Jordan CA (1989) Electroporation and electrofusion in cell biology. Plenum Press, New YorkGoogle Scholar
- Pavlin M, Flisar K, Kanduser M (2010) The role of electrophoresis in gene electrotransfer. J Memb Biol. doi: 10.1007/s00232-010-9276-z
- Valic B, Golzio M, Pavlin M, Schatz A, Faurie C, Gabriel B, Teissié J, Rols MP, Miklavcic D (2003) Effect of electric field induced transmembrane potential on spheroidal cells: theory and experiment. Biophys J 32:519–528Google Scholar