Cell–Cell Electrofusion: Optimization of Electric Field Amplitude and Hypotonic Treatment for Mouse Melanoma (B16-F1) and Chinese Hamster Ovary (CHO) Cells

Abstract

Efficient electroporation of cells in physical contact induces cell fusion, and this process is known as electrofusion. It has been shown that appropriate hypotonic treatment of cells before the application of electric pulses can cause a significant increase in electrofusion efficiency. First, the amplitudes of the electric field were determined spectrofluorometrically, where sufficient permeabilization in hypotonic buffer occurred for B16-F1 and CHO cells. In further electrofusion experiments 14 +/- 4% of fused cells for B16-F1 and 6 +/- 1% for CHO was achieved. These electrofusion efficiencies, determined by double staining and fluorescence microcopy, are comparable to those of other published studies. It was also confirmed that successful electroporation does not necessarily guarantee high electrofusion efficiency due to biological factors involved in the electrofusion process. Furthermore, not only the extension of electrofusion but also cell survival depends on the cell line used. Further studies are needed to improve overall cell survival after electroporation in hypotonic buffer, which was significantly reduced, especially for B16-F1 cells. Another contribution of this report is the description of a simple modification of the adherence method for formation of spontaneous cell contact, while cells preserve their spherical shape.

A high‐throughput dielectrophoresis‐based cell electrofusion microfluidic device

“…Moreover, the electric field generated between the plate electrodes is Ning Hu 1,2 Jun Yang 1 Zheng-Qin Yin 3 Ye Ai uniform, which leads to an equal probability of reversible electroporation and electrofusion of the aligned cells. Thus, the probability of unwanted multi-cell electrofusion in the conventional electrofusion device is relatively high [10].…”

Planning of Electroporation-Based Treatments Using Web-Based Treatment-Planning Software

“…The phenomenon is considered to be a universal method and platform technology because all types of cells (animal, plant and microorganisms) are affected by electroporation (Miklavčič 2012). In fact, many electroporation-based applications have already been identified and developed, such as electrochemotherapy (ECT) of tumors , nonthermal irreversible ablation of tumors Maor et al 2009), gene therapy (Heller and Heller 2010), food preservation (Toepfl et al 2007) and others (Daugimont et al 2010;Gusbeth et al 2009;Ušaj et al 2010).…”

Electroporation of adherent cells with low sample volumes on a microscope stage

“…The efficiency of electroporation with our new device (Fig.3D) is comparable to that of a normal volume electroporator that requires the adherent cells to be removed from the substratum (Usaj et al, 2010;Yumura et al, 1995).…”