dc.contributor.author | Vadeikienė, Roberta | |
dc.contributor.author | Jakštys, Baltramiejus | |
dc.contributor.author | Ugenskienė, Rasa | |
dc.contributor.author | Šatkauskas, Saulius | |
dc.contributor.author | Juozaitytė, Elona | |
dc.date.accessioned | 2023-09-18T16:35:43Z | |
dc.date.available | 2023-09-18T16:35:43Z | |
dc.date.issued | 2022 | |
dc.identifier.uri | https://etalpykla.vilniustech.lt/handle/123456789/115307 | |
dc.description.abstract | Non‐adherent cells are difficult to transfect with chemical‐mediated delivery methods. Electroporation is an attractive strategy to transfer the molecules of interest into suspension cells. Care must be taken with the viability of the transfected cells since parameters, which increase cell membrane permeability, subsequently increase transfection efficiency, leading to higher cell death indices. We intended to evaluate the distribution of hard‐to‐transfect UT‐7 cells among different subpopulations: transfected/viable, untransfected/viable, transfected/dead, and untransfected/dead populations, for a better understanding of the relation between gene electrotransfer efficacy and cell death. The following electroporation parameters were tested: pulse strength, duration, plasmid DNA concentration, and ZnSO4 as DNase inhibitor. BTX T820 square‐wave generator was used, and 48 h after electroporation, cells were observed for viability and fluorescence analysis. Increasing pulse strength correlated directly with an increased ratio of pEGFP‐positive cells and inversely with cell viability. The best results, representing 21% pEGFP positive/viable cells, were obtained after EP with 1 HV 1400 V/cm pulse of 250 μs duration using 200 μg/mL plasmid concentration. Results demonstrated that plasmid concentration played the most significant role in pEGFP electrotransfer into UT‐7 cells. These results can represent a relevant improvement of gene electrotransfer to obtain genetically modified suspension cells for further downstream experiments. | eng |
dc.format | PDF | |
dc.format.extent | p. 2672-2687 | |
dc.format.medium | tekstas / txt | |
dc.language.iso | eng | |
dc.relation.isreferencedby | Science Citation Index Expanded (Web of Science) | |
dc.relation.isreferencedby | Scopus | |
dc.relation.isreferencedby | DOAJ | |
dc.relation.isreferencedby | PubMed | |
dc.source.uri | https://doi.org/10.3390/biomedicines10112687 | |
dc.title | Systemic optimization of gene electrotransfer protocol using hard‐to‐transfect UT‐7 cell line as a model | |
dc.type | Straipsnis Web of Science DB / Article in Web of Science DB | |
dcterms.accessRights | This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/) | |
dcterms.license | Creative Commons – Attribution – 4.0 International | |
dcterms.references | 40 | |
dc.type.pubtype | S1 - Straipsnis Web of Science DB / Web of Science DB article | |
dc.contributor.institution | Lietuvos sveikatos mokslų universitetas | |
dc.contributor.institution | Vytauto Didžiojo universitetas | |
dc.subject.researchfield | N 010 - Biologija / Biology | |
dc.subject.researchfield | N 011 - Biofizika / Biophysics | |
dc.subject.studydirection | D01 - Biologija / Biology | |
dc.subject.en | electroporation | |
dc.subject.en | optimization | |
dc.subject.en | plasmid DNA transfer | |
dc.subject.en | gene electrotransfer (GET) | |
dc.subject.en | non‐adherent cells | |
dc.subject.en | UT‐7 cell line | |
dcterms.sourcetitle | Biomedicines | |
dc.description.issue | iss. 11 | |
dc.description.volume | vol. 10 | |
dc.publisher.name | MDPI | |
dc.publisher.city | Basel | |
dc.identifier.doi | 141740565 | |
dc.identifier.doi | 2-s2.0-85141798816 | |
dc.identifier.doi | 85141798816 | |
dc.identifier.doi | 1 | |
dc.identifier.doi | 000881049700001 | |
dc.identifier.doi | 10.3390/biomedicines10112687 | |
dc.identifier.elaba | 157547610 | |