Electric pulse-induced fusion of mouse lymphoma cells: Roles of divalent cations and membrane lipid domains

Ohno‐Shosaku, Takako; Okada, Yasunobu

doi:10.1007/bf01871522

Cited by 47 publications

(12 citation statements)

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“…Electrofusion yield is also improved by the presence of bivalent cations in the medium (Ohno-Shosaku and Okada 1985;Vienken and Zimmermann 1985), while the presence of monovalent ions decreased the fusion yield . Nevertheless, cell electrofusion is a complex process and several biologically active substances affect its yield (Grobner, Velizarov, Berg 1996;Velizarov and Berg 1998a;Velizarov et al 1998b;Liu et al 2000).…”

Section: Electroporation Medium Compositionmentioning

confidence: 99%

Electroporation in Biological Cell and Tissue: An Overview

Kandušer

Miklavčič²

2008

Electrotechnologies for Extraction From Food Plants and Biomaterials

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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.

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Section: Electroporation Medium Compositionmentioning

confidence: 99%

Electroporation in Biological Cell and Tissue: An Overview

Kandušer

Miklavčič²

2008

Electrotechnologies for Extraction From Food Plants and Biomaterials

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“…Furthermore, high cell density also reduces the electric conductivity of the suspension and may be partially responsible for the enhancement of the cell viability by reducing the pulse current. 18 To generalize the post-pulse pelleting technique, we tested four more cell lines, NK-L, K-562, L1210 and MC2 cells. We found the same protection effect of pellet on viability (Figure 4a, b and c).…”

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

confidence: 99%

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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“…The displaced Ca2+ may rebind to the plasma membrane and enhance stabilization of the membrane after the fusion event. Recently the absolute requirement of Ca2+ in the electrofusion of mouse lymphoma cells was also reported (17).…”

mentioning

confidence: 99%

“…The mechanisms involved in electrically induced cell fusion (electrofusion) have been discussed mainly in relation to the physical nature of the lipid domain of the plasma membrane (17,30), as described by Ahkong et al (3) in chemically induced cell fusion. However, there are some unique features to be considered in electrofusion.…”

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confidence: 99%