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

Masa Omerzel

Masa Omerzel

  • Department of Experimental Oncology, Institute of Oncology LjubljanaLjubljana, Slovenia
  • Faculty of Health Sciences, University of LjubljanaLjubljana, Slovenia
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Omerzel, Masa
, 
Bostjan Markelc

Bostjan Markelc

  • Department of Experimental Oncology, Institute of Oncology LjubljanaLjubljana, Slovenia
  • Biotechnical Faculty, University of LjubljanaLjubljana, Slovenia
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Markelc, Bostjan
, 
Simona Kranjc Brezar

Simona Kranjc Brezar

Department of Experimental Oncology, Institute of Oncology LjubljanaLjubljana, Slovenia
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Brezar, Simona Kranjc
, 
Gregor Sersa

Gregor Sersa

  • Department of Experimental Oncology, Institute of Oncology LjubljanaLjubljana, Slovenia
  • Faculty of Health Sciences, University of LjubljanaLjubljana, Slovenia
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Sersa, Gregor
 and 
Maja Cemazar

Maja Cemazar

  • Department of Experimental Oncology, Institute of Oncology LjubljanaLjubljana, Slovenia
  • Faculty of Health Sciences, University of PrimorskaIzola, Slovenia
  • University of LjubljanaSlovenia
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Cemazar, Maja
  
Apr 21, 2025
Efficient gene transfer by pulse parameters for electrochemotherapy of cells in vitro and in muscle and melanoma tumors in mice's Cover Image
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Article Category: research article
Published Online: Apr 21, 2025
Page range: 203 - 212
Received: Mar 24, 2025
Accepted: Apr 03, 2025
DOI: https://doi.org/10.2478/raon-2025-0027
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© 2025 Masa Omerzel et al., published by Sciendo
This work is licensed under the Creative Commons Attribution 4.0 International License.

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 pulses Pulse direction Frequency (Hz)
GET1 250 100 8 unipolar 1
GET2 300 100 8 unipolar 1
GET3 300 100 3 bipolar 1
GET4 300 100 8 unipolar 5000
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Journal Subjects:
Medicine, Clinical Medicine, Internal Medicine, Haematology, Oncology, Radiology
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