Articles Vol. 66 No. 3 26/05/2025

18. EVALUATING PROLIFERATION POTENTIAL OF CAR-T CELLS WHEN CO-CULTURED WITH CD19 CANCER CELL LINES IN VITRO

Hang Ngo Thu1,2, Mao Can Van1,2
1 Department of Pathophysiology, Vietnam Military Medical University
2 Bộ môn Sinh lý bệnh, Học viện Quân y
Corresponding author: canvanmao@vmmu.edu.vn
DOI: 10.52163/yhc.v66i3.2508
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Abstract

Objective: To evaluate the proliferation ability of CAR-T when co-cultured with CD19+-expressing cell lines in vitro.

Subjects and methods: Transfering gene into K562 cells using the Transposon-transposase vector system combined with electroporation. PBMC cells were eparated and analyzed PBMC by flow cytometry. PBMC were transposed gene using P3 Primary Cells transfection solution, 1 µg SB100X plasmid and 3 µg CD19RCD137/pSB plasmid. Co-culturing CAR-T and CD19+ cells at a ratio of 1:1.

Results: The results showed that on 4-day culture, both CAR-T CD19RCD137 and CAR-T iCasp9-IL15 cells had the ability to proliferate during co-culture with CD19+ cells. The number of CD19RCD137 CAR-T cells tended to decrease, but the number of 4th CAR-T generation still increased during co-cultured with K562 without expressing CD19. On 11-day  culture, there were almost no CD19RCD137 CAR-T cells and a significant decrease in the number of iCasp9-IL15 CAR-T cells.

Conclusion: CAR-T cells have the ability to proliferate strongly during co-culture with CD19+ cell lines but have no ability to proliferate during co-culture with CD19- cell lines.

Keywords
CAR-T cells CD19 CD19RCD137 and iCasp9-IL15.
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