Electroporation and Electrofusion

IRE has been studied extensively with in vitro cellular systems. IRE has also been studied as method to destroy prokaryotic (Sale and Hamilton, 1967) and eukaryotic cells in vitro and has gained momentum recently as a method to kill microorganisms (Vemhes et al., 

mammalian normal cells (Vernhes et al., 1999), and mammalian cancer cells (Miller et al., 2005) in vitro and tumors (Nuccitelli et al., 2006). These studies have demonstrated the ability of IRE to completely eradicate an entire population of cells in vitro without inducing any thermal damage. 

Lee et al. hypothesized that electrical injury is often characterized by the preferential death of large mammalian cells (skeletal muscle, nerves) in tissue regions where insignificant temperature rise occurs (Esser et al., 2007 Bhatt et al., 1990 Lee et al., 

With the key distinction between shock trauma and temperature change, this 

The electrical properties of any material, including biological tissue, can be broadly separated into two categories conducting and insulating. In a conductor, the electric charges move freely in response to the application of an electric field, whereas in an insulator (dielectric), the charges are fixed and not free to move. 

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Neumann E, Sowers AE, Jordan CA (1989) Electroporation and electrofusion in cell biology. Plemun Press, New York... 

Nickoloff, J.A. (Ed.) (1995). Methods in Molecular Biology, Vol. 48 Animal Cell Electroporation and Electrofusion Protocols. Human Press, pp. 3-38. 

Neumann, E., Sowers, A.E., and Jordon, C.A., Electroporation and Electrofusion in Cell Biology, Plenum Press, New York, 1989. 

The physical properhes of hpid bilayers are those that define their response to external perturbations. Knowing the mechanical and rheological characteristics of hpid membranes will prepare us to tackle problems related to stress induced in bilayers by electric fields and the phenomena that it triggers, for example, dynamics of vesicle and cell deformahon, bilayer instability, electroporation, and electrofusion. 

The phenomenon of membrane electrofusion is of particular interest, because of its widespread use in cell biology and biotechnology (e.g., [162-164] and the references cited therein). The application of electrofusion to cells can lead to the creation of multinucleated viable cells with new properties (this phenomenon is also known as hybridization) (e.g., [164]). In addition, electroporation and electrofusion are often used to introduce molecules like proteins, foreign genes (plasmids), antibodies, and drugs into cells. 

Chang, D.C. (1992) Structure and dynamics of electric field-induced membrane pores as revealed by rapid-freezing electron microscopy, in Guide to Electroporation and Electrofusion (eds D.C. Chang, B.M. Chassy, J.A. Saunders, and A.E. Sowers), Academic Press, San Diego, pp. 9-27. 

Use of a high field to activate a membrane enzyme was first reported by Witt et al. (25) in 1976. They used dc pulses of approximately 1 kV/cm and of 1-ms duration to induce ATP synthesis by the chloroplast ATPase. Following this initial work, there have been many reports on 1-kV/ cm dc field-induced ATP synthesis in different ATP synthetic systems (see the literature cited in references 13 and 14). The main conclusion from these studies is that an applied field-induced transmembrane potential can facilitate ATP release from the enzyme whether a PEF can affect enzyme turnover is not clear. Because 1-kV/ cm dc fields also cause severe Joule heating of a sample suspension, thermal effects cannot be easily avoided except when very short electric pulses (microseconds) are used. Thus, the method has limited utility for electroactivation experiments. The PEF method is, however, quite popular for the study of electroporation and electrofusion of cell membranes (see the chapter by J. Weaver in this volume), electroinsertion of membrane proteins (26), and electrotransfection of cells (27). 

Tsong, T. Y. Electroporation of cell membranes Its mechanisms and applications. In Electroporation and Electrofusion in Cell Biology Neumann, E. et al., Eds. Plenum New York pp 149-163. 

Animal Cell Electroporation and Electrofusion Protocols, edited by Jac A. Nickoloff. 1995 47. Electroporation Protocols for Microorganisms, edited by Jac A. Nickoloff 1995... 

At the cellular level, the cell membranes of polarized cells are of the order of 10 kV/mm, and additional field strengths may easily bring the membrane into a nonlinear region even without cell excitation. Cell excitation, the opening of membrane channels, and the creation of an action potential are the result of nonlinear processes. Electroporation and electrofusion of cells in vitro (Section 10.11) are also processes in the nonlinear region. 

Weaver, J.C., Barnett, A., 1992. Guide to Electroporation and Electrofusion in Progress toward a Theoretical Model of Electroporation Mechanism Membrane Electrical Behavior and Molecular Transport, 26. 

Techaumnat B, Washizu M (2007) Analysis of the effects of an orifice plate on the membrane potential in electroporation and electrofusion of cells. J Phys D ApplPhys 40 1831-1837... 

Neumann, E., Schaefer-Ridder, M., Wang, Y., and Hofschneider, P. H. (1982) Gene transfer into mouse myeloma cells by electroporation in high electric fields. EMBO J. 7, 841-845. Neumann, E., Sowers, A. E., and Jordon, C. A. (eds.) (1989) Electroporation and Electrofusion in Cell Biology. Plenum, New York. 

Chang DC, Chassy BM, Saunders JA, Sowers AE (1992) Guide to electroporation and electrofusion. Academic, San Diego... 

Crystallographic Methods and Protocols, edited by Christopher Jones, Barbara Mulloy, and Mark Sanderson, 1996 55. Plant Cell Electroporation and Electrofusion Protocols,... 

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