Control by pulse parameters of electric field-mediated gene transfer in mammalian cells

Wolf, Hendrik; Rols, Marie‐Pierre; Boldt, Elvira; Neumann, Eberhard; Teissié, Justin

doi:10.1016/s0006-3495(94)80805-7

Cited by 208 publications

(188 citation statements)

References 44 publications

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“…There was an apparent sharp maximum for marker expression at 25 gg of plasmid DNA per tissue injection. This un-10,000 expected dose dependence in vivo is different from in vitro electroporation, in which the DNA dose dependence exhibits saturation [15]. One important advantage of electroporation in vitro is the high efficiency of intracellular DNA delivery.…”

Section: Resultsmentioning

confidence: 94%

In vivo gene electroinjection and expression in rat liver

et al. 1996

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In vivo targeted gene transfer by non-viral vectors is subjected to anatomical constraints depending on the route of administration. Transfection efficiency and gene expression in vivo using non-viral vectors is also relatively low. We report that in vivo electropermeabilization of the liver tissue of rats in the presence of genes encoding luciferase or ~-galactosidase resulted in the strong expression of these genetic markers in rat liver cells. About 31)-40% of the rat liver cells electroporated expressed the ~galactosidase genetic marker 48 h after electroporation. The marker expression was also detected at least 21 days after transfection at about 5% of the level 48 h after electroporation. The results indicate that gene transfer by electroporatinn in vivo may avoid anatomical constraints and low transfection efficiency.

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Section: Resultsmentioning

confidence: 94%

In vivo gene electroinjection and expression in rat liver

et al. 1996

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“…On the other hand, the post-pulse pelleting technique should be good for loading charged molecules, such as DNA, since it is known that electrophoresis of DNA during pulses is the major mechanism for electrotransfection of cells. 7,11 Centrifugation immediately after pulses is not expected to significantly reduce the efficiency of DNA internalization. 20 With improved cell viability in the post-pulse centrifugation pellet, the pelleting technique should greatly enhance the electrotransfection efficiency.…”

Section: ) the Other For Suspension Incubation ( ̅ ᭺) (D) The Dementioning

confidence: 99%

“…4,5 Also, the parameters in electroporation are easier to control and the efficiency in some cases is higher than those by other methods. [6][7][8][9] Moreover, it avoids biological contamination that potentially exists in virus-induced methods. It offers an attractive way for ex vivo gene delivery if the transfection efficiency can be significantly improved.…”

Section: Introductionmentioning

confidence: 99%

“…10,11 The second step is to allow cell membranes to recover to their original impermeable state. 7 Initially, in the reversible breakdown step, the higher the pulse energy (including pulse duration and pulse strength), the higher the intake by electroloading. 12 If the pulse electrical field is too strong, an irreversible breakdown occurs, and the efficiency of electrotransfection and electroloading suffers due to the drop of cell viability.…”

Section: Introductionmentioning

confidence: 99%

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Improving electrotransfection efficiency by post-pulse centrifugation

Ross

Hui

1999

Gene Ther

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We have demonstrated that the viability of electrodorf desktop centrifuge. Pelleting improves the cell viability transfected adherent CHO and suspended NK-L, K-562, over the whole range of the NK-L, K-562, L1210 and MC2 L1210 and MC2 cells is improved if pelleting by centrifugcell concentrations studied. When this pelleting method is ation is performed immediately after pulsing. The protecapplied to load CHO cells with FITC-dextran (41 000 MW), tion effect on cell viability is cell line-and pellet thicknessnot only is the success rate close to 100%, but the growth dependent. For forming CHO cell pellets, centrifugation rate is similar to the control, which is far better than the force (300-13 000 g) and duration are not crucial; about conventional electroporation method. Furthermore, the five to 10 cell layers in the pellet provide the optimal protectransfection efficiency of the five cell lines in pellet is sigtion effect. NK-L, K-562, L1210 and MC2 cell pellets are nificantly higher than that in suspension. optimally formed by centrifugation at 13 000 g in an Eppen-

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“…Induced transmembrane potential depends on cell and tissue parameters (tissue conductivity, cell size, shape, and distribution) 12,20,22 and pulse parameters (pulse duration, amplitude, and number of pulses). 24 After treatment, cell membrane reseals provided the applied voltage was, although high enough to cause cell membrane permeabilization, still low enough not to cause irreversible permeabilization, hence permanent damage.…”

Section: Introductionmentioning

confidence: 99%

A Numerical Model of Skin Electropermeabilization Based on In Vivo Experiments

2007

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Abstract-As an alternative to viral methods that are controversial because of their safety issues, chemical and physical methods have been developed to enhance gene expression in tissues. Reversible increase of the cell membrane permeability caused by the electric field-electroporation-is currently one of the most efficient and simple nonviral methods of gene transfer. We performed a series of in vivo experiments, delivering plasmids to rat skin using external plate electrodes. The experiments showed that skin layers below stratum corneum can be permeabilized in this way. In order to study the course of skin tissue permeabilization by means of electric pulses, a numerical model using the finite element method was made. The model is based on the tissue-electrode geometry and electric pulses used in our in vivo experiments. We took into account the layered structure of skin and changes of its bulk electrical properties during electroporation, as observed in the in vivo experiments. We were using tissue conductivity values found in literature and experimentally determined electric field threshold values needed for tissue permeabilization. The results obtained with the model are in good agreement with the in vivo results of gene transfection in rat skin. With the model presented we used the available data to explain the mechanism of the tissue electropermeabilization propagation beyond the initial conditions dictated by the tissue initial conductivities, thus contributing to a more in-depth understanding of this process. Such a model can be used to optimize and develop electrodes and pulse parameters.

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Control by pulse parameters of electric field-mediated gene transfer in mammalian cells

Cited by 208 publications

References 44 publications

In vivo gene electroinjection and expression in rat liver

In vivo gene electroinjection and expression in rat liver

Improving electrotransfection efficiency by post-pulse centrifugation

A Numerical Model of Skin Electropermeabilization Based on In Vivo Experiments

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