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Date Published: 28/06/2025
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DOI: https://doi.org/10.56535/jmpm.v50i5.1273
Issue
Vol. 50 No. 5 (2025): TẠP CHÍ Y DƯỢC HỌC QUÂN SỰ SỐ 5 - 2025
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Articles
How to Cite
Cấn, V. M., & Ngo, T. H. (2025). Transposition of Sleeping beauty plasmid and minicircle into T cells using Nucleofector technology. Journal of Military Pharmaco-medicine, 50(5), 52-59. https://doi.org/10.56535/jmpm.v50i5.1273

Transposition of Sleeping beauty plasmid and minicircle into T cells using Nucleofector technology

Cấn Văn Mão1, , Ngo Thu Hang1
1 Bộ môn Sinh lý bệnh, Học viện Quân y

Main Article Content

Abstract

Objective: Study on the transposition of Sleeping Beauty plasmid and minicircle in the transposition of chimeric antigen receptors into T cells using Nucleofector technology. Subjects and methods: Fresh blood samples were processed and separated peripheral blood mononuclear cells (PBMC), then this cells were counted and analyzed by flow cytometry, the transpositions of plasmid and minicircle were conduct to create CAR-T. Finally, the transposition efficiency was evaluated by flow cytometry analysis. Results: The transposition efficiency of the second generation CAR-T CD19RCD137/pSB plasmid construct (25.88%) was higher than that of the fourth-generation iCasp9-IL15/pSB CAR-T (14.72%), and the minicircle CAR-T CD19RCD137/pSB (35.12%) was higher than that of the iCasp9-IL15/pSB CAR-T (23.48%) with p<0.05. The viable cell rate of the second-generation CD19RCD137/pSB CAR-T (48.9%) was higher than that of the fourth-generation iCasp9-IL15/pSB CAR-T (25.4%), and the minicircle CD19RCD137/pSB CAR-T (35.12%) was higher than that of the iCasp9-IL15/pSB CAR-T (23.48%) with p<0.05. Cell proliferation results (4 days) with IL-2 (100U/mL) demonstrated that the PBMC population retained its function after the transposition. Conclusion: Transposition of CAR-T cell blocks based on the second- and fourth-generation plasmid and minicircle were successfully conducted that created a foundation for proliferation experiments and testing T cell function.

Article Details

Keywords

Sleeping Beauty plasmid, Sleeping Beauty minicircle, Nucleofector technology

References

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