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STUDY ON THE PROCESS OF CAR-T WITH AAPC CELL CO-CULTURE TO OPTIMIZE QUALITY OF CAR-T CELLS FOR THE TREATMENT OF ACUTE LYMPHOBLASTIC LEUKEMIA
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Abstract
Objective: To optimize the co-culture process of CAR-T and aAPC 1D2 cells in order to obtain CAR-T cells with the highest yield and quality. Subjects and Methods: PBMCs were transposed using the 4D Amaxa Nucleofector device with the EO-115 program. CAR-T cells were analyzed by flow cytometry on the BD FACSLyric system after each transposition. Subsequently, CAR-T cells were co-cultured with aAPCs in CTS Optimizer T-cell expansion medium. aAPC cells were added to the CAR-T culture medium at ratios equivalent to 1X or 2X live CAR-T cells. Results: When co-cultured with aAPC 1D2 at ratios of 1:2 and 1:5, CAR-T cells proliferated efficiently and comparably. The CAR-T cell population reached 52.63% CAR expression on day 14 and 81.13% on day 21. Conclusion: The optimal co-culture ratio was determined to be CAR-T: aAPC 1D2 at a 1:2 ratio in the volume. A minimum of three transpositions (with aAPC 1D2 added every 7 days) was required to obtain CAR-T cells with robust antigen expression.
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Keywords
Artificial antigen-presenting cells, culture and transposition.
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