Dielectrophoretic Manipulation of Cell Transfection Efficiency during Electroporation Using a Center Needle Electrode

Abstract

Long duration electric pulses are frequently used to facilitate DNA electrotransfer into cells and tissues, while electroporation pulses can be combined with electrophoresis to maximize the transfection efficiency. In this work, we present the dielectrophoresis (DEP)-assisted methodology for electrotransfer of plasmid DNA (3.5 kbp pmaxGFP) into mammalian cells (CHO-K1). A prototype of an electroporation cuvette with center needle electrode for DEP-assisted transfection is presented resulting in a 1.4-fold of transfection efficiency increase compared to the electroporation-only procedure (1.4 kV/cm × 100 µs × 8). The efficiency of transfection has been compared between three DEP frequencies of 1, 100, and 1 MHz. Lastly, the effects of exposure time (1, 3, and 5 min) during the DEP application step have been determined. It is concluded that the proposed methodology and exposure setup allow a significant improvement of transfection efficiency and could be used as an alternative to the currently popular electrotransfection techniques.

Effect of Experimental Electrical and Biological Parameters on Gene Transfer by Electroporation: A Systematic Review and Meta-Analysis

“…Finally, data from 228 eligible papers (Figure 2) were extracted by two investigators [6,13,31,33,[37][38][39] (Supplementary List S1). Of the 228 eligible papers, only 35 papers were appropriate based on the risk of bias assessment [31,37,38,47,48,82,94,105,116,127,130,131,[138][139][140][141][144][145][146]157,158,169,173,176,177,[186][187][188][189][190]196,199,214,238,253]. For the meta-analysis, data of standard deviation or error and sample size were additionally needed.…”

“…For the meta-analysis, data of standard deviation or error and sample size were additionally needed. Of 35 eligible papers based on the risk of bias, standard deviation or error and sample size were only reported in 23 papers [31,37,38,47,48,116,[139][140][141]145,146,157,158,169,173,176,177,187,188,190,196,199,253] (Figure 2). The first search in August 2018 retrieved 6133 papers.…”

Transfection by Electroporation of Cancer and Primary Cells Using Nanosecond and Microsecond Electric Fields

“…In a variety of studies, micro-or nanosecond pulses were applied to widely used mammalian hosts-Chinese hamster ovary (CHO) cells [8][9][10][14][15][16], and to evaluate the efficacy of a gene electrotransfection, different protein-encoding plasmids were used. However, one of the most commonly used plasmids are green fluorescent protein (GFP)-encoding plasmids (3.5 kb, pMAX-GFP [12,14,[16][17][18]; 4.7 kb, pEGFP-N1 [14,15]) and luciferase-expressing plasmids (4.6 kbp pGL4.13 [16,19]). The use of model cell lines such as CHO and standard plasmids is a common practice to improve the repeatability of the results during derivation of the optimal electroporation conditions.…”