Ascorbate Increases Electrotransformation Efficiency of Intact Maize Cells

Sabri, N.; Pelissier, B.; Teissié, Justin

doi:10.1006/abio.1998.2878

Cited by 12 publications

(7 citation statements)

References 18 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Reactive oxidative species are produced in response to oxidative stress in mammalian and plant cells exposed to electric pulses (Biedinger et al 1990;Teissié 1995a, Maccarrone et al 1995;Sabri et al 1996bSabri et al , 1998. Even if the cell viability is not directly correlated with reactive oxidative species production, gene electrotransfer efficiency is improved by post-pulse treatment with antioxidants, which protect the cell from reactive oxidative species (Sabri et al 1996;Sabri et al 1998).…”

Section: Transfection Of Intact Plant Tissuementioning

confidence: 99%

See 1 more Smart Citation

Electroporation in Biological Cell and Tissue: An Overview

Kandušer

Miklavčič²

2008

Electrotechnologies for Extraction From Food Plants and Biomaterials

View full text Add to dashboard Cite

In this chapter, basics and mechanisms of electroporation are presented. Most important electric pulse parameters for electroporation efficiency for different applications that involve introduction of small molecules and macromolecules into the cell or cell membrane electrofusion are described. In all these applications, cell viability has to be preserved. However, in some biotechnological applications, such as liquid food sterilization or water treatment, electroporation is used as a method for efficient cell killing. For all the applications mentioned above, besides electric pulse parameters, other factors, such as electroporation medium composition and osmotic pressure, play significant roles in electroporation effectiveness. For controlled use of the method in all applications, the basic mechanisms of electroporation need to be known. The phenomenon was studied from the single-cell level and dense cell suspension that represents a simplified homogenous tissue model, to complex biological tissues. In the latter, different cell types and electric conductivity that change during the course of electric pulse application can significantly affect the effectiveness of the treatment. For such a complex situation, the design and use of suitable electrodes and theoretical modeling of electric field distribution within the tissue are essential. Electroporation as a universal method applicable to different cell types is used for different purposes. In medicine it is used for electrochemotherapy and genetherapy. In biotechnology it is used for water and liquid food sterilization and for transfection of bacteria, yeast, plant protoplast, and intact plant tissue. Understanding the phenomenon of electroporation, its mechanisms and optimization of all the parameters that affect electroporation is a prerequisite for successful treatment. In addition to the parameters mentioned above, different biological characteristics of treated cell affect the outcome of the treatment. Electroporation, gene electrotransfer and electrofusion are affected by cell membrane fluidity, cytoskeleton, and the presence of the cell wall in bacteria yeast and plant cells. Thus, electroporation parameters need to be specifically optimized for different cell types.

show abstract

Section: Transfection Of Intact Plant Tissuementioning

confidence: 99%

“…Even if the cell viability is not directly correlated with reactive oxidative species production, gene electrotransfer efficiency is improved by post-pulse treatment with antioxidants, which protect the cell from reactive oxidative species (Sabri et al 1996;Sabri et al 1998).…”

Section: Transfection Of Intact Plant Tissuementioning

confidence: 99%

Electroporation in Biological Cell and Tissue: An Overview

Kandušer

Miklavčič²

2008

Electrotechnologies for Extraction From Food Plants and Biomaterials

View full text Add to dashboard Cite

show abstract

“…The ROS generated due to EP on the cell surface must affect the plasmid DNA very rapidly, since there was no increase in transfection efficiency if tempol was added after the EP. A positive effect of antioxidants, such as ascorbic acid, on expression levels after electrotransfection has been reported in plant cells, 47 but to our best knowledge, there has been no report of increased electrotransfection efficiency in animal cells in vitro or in vivo following (due to) the addition of antioxidants.…”

Section: Discussionmentioning

confidence: 94%

Muscle gene electrotransfer is increased by the antioxidant tempol in mice

et al. 2011

View full text Add to dashboard Cite

Electropermeabilization (EP) is an effective method of gene transfer into different tissues. During EP, reactive oxygen species (ROS) are formed, which could affect transfection efficiency. The role of generated ROS and the role of antioxidants in electrotransfer in myoblasts in vitro and in Musculus tibialis cranialis in mice were, therefore, investigated. We demonstrate in the study that during EP of C2C12 myoblasts, ROS are generated on the surface of the cells, which do not induce long-term genomic DNA damage. Plasmid DNA for transfection (pEGFP-N1), which is present outside the cells during EP, neutralizes the generated ROS. The ROS generation is proportional to the amplitude of the electric pulses and can be scavenged by antioxidants, such as vitamin C or tempol. When antioxidants were used during gene electrotransfer, the transfection efficiency of C2C12 myoblasts was statistically significantly increased 1.6-fold with tempol. Also in vivo, the transfection efficiency of M. tibialis cranialis in mice was statistically significantly increased 1.4-fold by tempol. The study indicates that ROS are generated on cells during EP and can be scavenged by antioxidants. Specifically, tempol can be used to improve gene electrotransfer into the muscle and possibly also to other tissues.

show abstract

“…This can brings a lethal effect on the cells [72]. But plasmid DNA was another target of ROS as shown by the protective effect on electrotransfection of antioxydants [73]. Nevertheless, co-injection of a coblockpolymer known to protect tissue against burns did not induce any protection [68].…”

Section: Tissue Damagementioning

confidence: 96%

Electrically-Assisted Nucleic Acids Delivery to Tissues In Vivo: Where Do We Stand?

Čemažar¹,

Golzio²,

Serša³

et al. 2006

CPD

Self Cite

View full text Add to dashboard Cite

Electropulsation (electroporation) is a physical method for delivery of various molecules into the cells in vitro and in vivo. It is an expanding field due to its applicability in cancer therapy, where combined application of electric pulses and chemotherapeutic drugs is used for treatment of cutaneous and subcutaneous nodules of different malignancies. Another application of electropulsation in vivo is electrogene therapy, where after injection of naked plasmid DNA and delivery of electric pulses directly to the tissue the expression of gene of interest can be obtained. However, the transfection efficiency of this methodology in vivo is still lower than with viral vectors. Nevertheless, due to the lack of immunogenicity of the method, easiness of the preparation of large quantities of endotoxin free plasmid DNA, control and reproducibility of the method and the development of electropulsators approved for the clinical use, electrically-assisted nucleic-acid delivery holds a great potential for the clinical application. This aim of this minireview is to critically discuss the main limitations and obstacles associated with electrogene therapy and the failures and problems as well as the successes. Topics on electric field distribution in the tissue, electrode geometries, construction of plasmid, modulation of extracellular space, tissue damage, pro-inflammatory and immune response as well as blood flow modification associated with application of electric pulses and injection of naked DNA are presented with possible directions how to overcome these limitations. Furthermore, for successful electrogene therapy in clinical setting it is of utmost importance to elucidate the mechanisms of DNA transfer into the cells of tissues in vivo. This will enable appropriate selection of electric pulse parameters and plasmid DNA constructs for each particular intended use. In the long run, this review should encourage other scientists to consider electrically assisted gene delivery for gene therapy as it matures.

show abstract

Ascorbate Increases Electrotransformation Efficiency of Intact Maize Cells

Cited by 12 publications

References 18 publications

Electroporation in Biological Cell and Tissue: An Overview

Electroporation in Biological Cell and Tissue: An Overview

Muscle gene electrotransfer is increased by the antioxidant tempol in mice

Electrically-Assisted Nucleic Acids Delivery to Tissues In Vivo: Where Do We Stand?

Contact Info

Product

Resources

About