Use this url to cite publication: https://hdl.handle.net/20.500.12512/116076
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Systemic Optimization of Gene Electrotransfer Protocol Using Hard-to-Transfect UT-7 Cell Line as a Model / Roberta Vadeikienė, Baltramiejus Jakštys, Rasa Ugenskienė, Saulius Šatkauskas, Elona Juozaitytė
Type of publication
Straipsnis Web of Science ir Scopus duomenų bazėje / Article in Web of Science and Scopus database (S1)
Author(s)
Jakštys, Baltramiejus | Vytautas Magnus University | |
Šatkauskas, Saulius | Vytautas Magnus University | |
Title
Systemic Optimization of Gene Electrotransfer Protocol Using Hard-to-Transfect UT-7 Cell Line as a Model / Roberta Vadeikienė, Baltramiejus Jakštys, Rasa Ugenskienė, Saulius Šatkauskas, Elona Juozaitytė
Publisher (trusted)
MDPI |
Is Referenced by
Date Issued
Date Issued |
---|
2022-10-24 |
Extent
p. 1-14.
Is part of
Biomedicines. Basel : MDPI, 2022, vol. 10, no. 11.
Version
Originalus / Original
Description
Article No. 2687.
elaba:157547610
OA, (CC BY) license.
Field of Science
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.
Type of document
type::text::journal::journal article::research article
ISSN (of the container)
2227-9059
WOS
000881049700001
Other Identifier(s)
(LSMU ALMA)991679987307106
Coverage Spatial
Šveicarija / Switzerland (CH)
Language
Anglų / English (en)
Bibliographic Details
40
Access Rights
Prieiga intranete / Intranet Access
File(s)biomedicines-10-02687-v2.pdf (2.13 MB) Intranet Access