Investigation of Plasmid DNA Delivery and Cell Viability Dynamics for Optimal Cell Electrotransfection In Vitro

Abstract

Electroporation is an effective method for delivering plasmid DNA molecules into cells. The efficiency of gene electrotransfer depends on several factors. To achieve high transfection efficiency while maintaining cell viability is a tedious task in electroporation. Here, we present a combined study in which the dynamics of both evaluation types of transfection efficiency and the cell viability were evaluated in dependence of plasmid concentration as well as at the different number of high voltage (HV) electric pulses. The results of this study reveal a quantitative sigmoidal (R2 > 0.95) dependence of the transfection efficiency and cell viability on the distance between the cell membrane and the nearest plasmid. We propose this distance value as a new, more accurate output parameter that could be used in further optimization studies as a predictor and a measure of electrotransfection efficiency.

Low-dose Polyethylenimine-Mediated Transfection to Generate Human Induced Pluripotent Stem Cell: an optimized protocol

“…Higher concentrations of pDNA require a higher amount of PEI for modi cation and permeabilization of endosomal membrane, which causes a higher cytotoxic effect of polyplexes on cell viability [42,43]. In addition, regardless of PEI dosage, an excess of pDNA itself can also contribute to cellular toxicity [44,45]. There are pieces of evidence showing that regardless of the transfection method, the concentration of nucleic acid is also critical for enhancing e ciency and minimizing toxicity [44,46].…”

Transfection types, methods and strategies: a technical review

“…For instance, the use of fetal bovine serum was shown to yield better transduction efficiency than bovine calf serum during transduction (Denning et al, 2013). (Haas et al, 2000;Ikeda et al, 2002;Denning et al, 2013) Potter & Heller, 2003;Molnar et al, 2004;Yao et al, 2009;Hornstein et al, 2016;Chopra et al, 2020;Sherba et al, 2020) (Stevenson et al, 2006;Terakawa et al, 2006;Tsen et al, 2009) (Wells et al, 1998;Dahlhoff et al, 2012;Mellott, Forrest & Detamore, 2013;Chow et al, 2016) (Ogawa, Tachibana & Kondo, 2006;Kinoshita & Hynynen, 2007;Zhou et al, 2012) (Mykhaylyk et al, 2007;Fouriki et al, 2010;Kardos & Rabussay, 2012;Wang et al, 2014) Likewise, polycations such as DEAE-dextran were shown to minimize repulsion forces between negatively-charged cells and facilitated viral transduction (Denning et al, 2013).…”

“…Prolonged electroporation with high voltage applied could potentially lead to cell damage and reduce transfection efficiency (Molnar et al, 2004). The electro-transfection efficiency can also be improved by increasing the number of electric pulses, but this may reduce the cell viability (Chopra et al, 2020). On the other side, the electro-transfection efficiency is dependent on the type of cell used and the electroporation condition should be optimized whenever a new cell type is going to be electro-transfected (Potter & Heller, 2003).…”

“…However, not only did efficiency not improve, cell survival also declined post-electroporation with further increment of plasmid concentration to 0.3 mg/ml (Yao et al, 2009). In circumstances when low electric pulse number is used, a high plasmid DNA concentration can be applied to compensate the low number of electric pulses to achieve a good electro-transfection efficiency (Chopra et al, 2020).…”

Peer Review #1 of "Transfection types, methods and strategies: a technical review (v0.1)"

“…Prolonged electroporation with high voltage applied could potentially lead to cell damage and reduce transfection efficiency (Molnar et al 2004). The electro-transfection efficiency can also be improved by increasing the number of electric pulses, but this may reduce the cell viability (Chopra et al 2020). On the other side, the electro-transfection efficiency is dependent on the type of cell used and the electroporation condition should be optimized whenever a new cell type is going to be electro-transfected (Potter and Heller 2003).…”