Feasibility study for the use of gene electrotransfer and cell electrofusion as a single-step technique for the generation of activated cancer cell vaccines

Ušaj, Marko (author), Pavlin, Mojca (author), Kandušer, Maša (author)

Cell-based therapies hold great potential for cancer immunotherapy. This approach is based on manipulation of dendritic cells to activate immune system against specific cancer antigens. For the development of an effective cell vaccine platform, gene transfer, and cell fusion have been used for modification of dendritic or tumor cells to express immune (co)stimulatory signals and to load dendritic cells with tumor antigens. Both, gene transfer and cell fusion can be achieved by single technique, a cell membrane electroporation. The cell membrane exposed to external electric field becomes temporarily permeable, enabling introduction of genetic material, and also fusogenic, enabling the fusion of cells in the close contact. We tested the feasability of combining gene electrotransfer and electrofusion into a single-step technique and evaluated the effects of electroporation buffer, pulse parameters, and cell membrane fluidity for single or combined method of gene delivery or cell fusdion. We determined the percentage of fused cells expressing green fluorescence protein (GFP) in a murine cell model of melanoma B16F1, cell line used in our previous studies. Our results suggest that gene electrotransfer and cell electrofusion can be applied in a single step. The percentage of viable hybrid cells expressing GFP depends on electric pulse parameters and the composition of the electroporation buffer. Furthermore, our results suggest that cell membrane fluidity is not related to the efficiency of the gene electrotransfer and electrofusion. The protocol is compatible with microfluidic devices, however further optimization of electric pulse parameters and buffers is still needed.

• Language: English
• Keywords: cancer cell vaccines, cell membrane fluidity, electrofusion, electroporation, gene electrotransfer, cell membrane fuidity
• Work type: Article
• Typology: 1.01 - Original Scientific Article
• Organization: MF - Faculty of Medicine; FE - Faculty of Electrical Engineering; FFA - Faculty of Pharmacy
• Publication status: Published
• Publication version: Version of Record
• Year: 2024
• Number of pages: Str. 377–389
• Numbering: Vol. 257, iss. 5/6
• PID: 20.500.12556/RUL-165160
• UDC: 616-085.37:616-006
• ISSN on article: 1432-1424
• DOI: 10.1007/s00232-024-00320-5
• COBISS.SI-ID: 204778499
• Publication date in RUL: 25.11.2024

Record is a part of a journal

• Title: The journal of membrane biology
• Shortened title: J. membr. biol.
• Publisher: Springer Nature
• ISSN: 1432-1424
• COBISS.SI-ID: 512024345

Licences

• License: CC BY 4.0, Creative Commons Attribution 4.0 International
• Link: http://creativecommons.org/licenses/by/4.0/
• Description: This is the standard Creative Commons license that gives others maximum freedom to do what they want with the work as long as they credit the author.

Secondary language

• Language: Slovenian
• Keywords: cepiva proti rakavim celicam, pretočnost celične membrane, elektrofuzija, elektroporacija, genski elektrotransfer, rak, imunoterapija, cepiva, celična membrana