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Efficient gene transfer by pulse parameters for electrochemotherapy of cells in vitro and in muscle and melanoma tumors in mice Cover

Efficient gene transfer by pulse parameters for electrochemotherapy of cells in vitro and in muscle and melanoma tumors in mice

Radiology and Oncology
Volume 59 (2025): Issue 2 (June 2025)
By: Masa Omerzel,  Bostjan Markelc,  Simona Kranjc Brezar,  Gregor Sersa and  Maja Cemazar  
Open Access
|Apr 2025

Figures & Tables

FIGURE 1.

The transfection efficiency timeline of different cell lines according to the gene electrotransfer (GET) protocol. Percent of transfected B16F10 (A), C2C12 (B) and L929 (C) cells and fluorescence intensity of B16F10 (D), C2C12 (E) and L929 (F) cells. *p < 0.05, statistically significant difference in GET1/GET2 compared with GET4 (A); *p < 0.05, statistically significant difference in GET1/GET4 compared with GET3 and GET2 (B); **p < 0.05, statistically significant difference in GET2 compared with GET3 (B); *p < 0.05, statistically significant difference in GET1/GET2/GET4 compared with GET4 (D and E)
The transfection efficiency timeline of different cell lines according to the gene electrotransfer (GET) protocol. Percent of transfected B16F10 (A), C2C12 (B) and L929 (C) cells and fluorescence intensity of B16F10 (D), C2C12 (E) and L929 (F) cells. *p < 0.05, statistically significant difference in GET1/GET2 compared with GET4 (A); *p < 0.05, statistically significant difference in GET1/GET4 compared with GET3 and GET2 (B); **p < 0.05, statistically significant difference in GET2 compared with GET3 (B); *p < 0.05, statistically significant difference in GET1/GET2/GET4 compared with GET4 (D and E)

FIGURE 2.

Transfection efficiency of different pulse parameter protocols in three different cell lines. Percent of transfected tumor (B16F10) and normal (C2C12, L929) cells after GET1 (A), GET2 (B), GET3 (C) and GET4 (D) treatment. *p < 0.05 indicates a statistically significant difference; GET = gene electrotransfer
Transfection efficiency of different pulse parameter protocols in three different cell lines. Percent of transfected tumor (B16F10) and normal (C2C12, L929) cells after GET1 (A), GET2 (B), GET3 (C) and GET4 (D) treatment. *p < 0.05 indicates a statistically significant difference; GET = gene electrotransfer

FIGURE 3.

Cell survival after different pulse parameters and gene electrotransfer (GET) in B16F10 (A), C2C12 (B) and L929 (C) cells. * = p < 0.05 indicates a statistically significant difference; EP = electroporation only; pGFP = plasmid only, which encodes the green fluorescent protein
Cell survival after different pulse parameters and gene electrotransfer (GET) in B16F10 (A), C2C12 (B) and L929 (C) cells. * = p < 0.05 indicates a statistically significant difference; EP = electroporation only; pGFP = plasmid only, which encodes the green fluorescent protein

FIGURE 4.

Transfection of B16F10 tumors. Untreated control tumors (A), GET2 (B) and GET4 (C) tumors. Nuclei are stained blue with Hoechst. The transfected cells are presented in green, indicating transfection with the GFP plasmid. Scale bar = 1000 μm
Transfection of B16F10 tumors. Untreated control tumors (A), GET2 (B) and GET4 (C) tumors. Nuclei are stained blue with Hoechst. The transfected cells are presented in green, indicating transfection with the GFP plasmid. Scale bar = 1000 μm

FIGURE 5.

Transfection of muscle. Control (A), GET2 (B), GET4 (C) and 1HV-4LV (D). The nuclei are stained blue with Hoechst. The transfected cells are presented in green, indicating transfection with the GFP plasmid. Scale bar = 1000 μm.
Transfection of muscle. Control (A), GET2 (B), GET4 (C) and 1HV-4LV (D). The nuclei are stained blue with Hoechst. The transfected cells are presented in green, indicating transfection with the GFP plasmid. Scale bar = 1000 μm.

Pulse parameters for different gene electrotransfer (GET) protocols for in vitro cell transfection

Experimental groups (protocol name)Voltage (V)Pulse duration (μs)No. of pulsesPulse directionFrequency (Hz)
GET12501008unipolar1
GET23001008unipolar1
GET33001003bipolar1
GET43001008unipolar5000
DOI: https://doi.org/10.2478/raon-2025-0027 | Journal eISSN: 1581-3207 | Journal ISSN: 1318-2099
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Language: English
Page range: 203 - 212
Submitted on: Mar 24, 2025
Accepted on: Apr 3, 2025
Published on: Apr 21, 2025
Published by: Association of Radiology and Oncology
In partnership with: Paradigm Publishing Services
Publication frequency: 4 issues per year
Keywords:
gene electrotransfer,
electrochemotherapy,
cells,
tumor,
muscle
Related subjects:
Medicine,
Clinical medicine,
Internal medicine,
Haematology, oncology,
Radiology

© 2025 Masa Omerzel, Bostjan Markelc, Simona Kranjc Brezar, Gregor Sersa, Maja Cemazar, published by Association of Radiology and Oncology
This work is licensed under the Creative Commons Attribution 4.0 License.

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