Presentation Information
[P03-415]Development of a Serum-Free Culture System with Five Kinds of Interleukin Cytokines and Insulin for T Cell Expansion
○Chao-Ling Yao1 (1. National Cheng Kung University (Taiwan))
Keywords:
T cells,Medium optimization,Insulin,Interleukin,Serum-free
[Purpose] Adoptive T cell therapy (ACT) requires the robust ex vivo expansion of functional T cells, but serum-containing culture systems, although supportive of proliferation, remain unsuitable for standardized clinical manufacturing. This study established a defined, serum-free culture platform that promoted effective CD8 T-cell proliferation.
[Method] CD3 T cells were purified from mononuclear cells and cultured under various cytokine and medium conditions. Using fractional factorial screening followed by steepest ascent optimization, we identified an optimal 5-cytokine cocktail (interleukin [IL]-2, IL-4, IL-7, IL-10, and IL-15), which markedly enhanced T-cell proliferation and increased the proportion of CD8 T cells.
[Results] Evaluation of basal media under serum-free conditions revealed that X-VIVO 15 effectively supported T-cell expansion in the presence of the optimal 5-cytokine cocktail. When insulin was added to X-VIVO 15 together with the optimal 5-cytokine cocktail, CD8 T-cell proliferation reached levels comparable to those achieved in 10% fetal bovine serum cultures while maintaining serum-free conditions.
[Conclusion] This optimized system provides a reproducible and scalable platform for serum-free expansion of human T cells, supporting the manufacturing of ACT and chimeric antigen receptor T cell therapies.
[Method] CD3 T cells were purified from mononuclear cells and cultured under various cytokine and medium conditions. Using fractional factorial screening followed by steepest ascent optimization, we identified an optimal 5-cytokine cocktail (interleukin [IL]-2, IL-4, IL-7, IL-10, and IL-15), which markedly enhanced T-cell proliferation and increased the proportion of CD8 T cells.
[Results] Evaluation of basal media under serum-free conditions revealed that X-VIVO 15 effectively supported T-cell expansion in the presence of the optimal 5-cytokine cocktail. When insulin was added to X-VIVO 15 together with the optimal 5-cytokine cocktail, CD8 T-cell proliferation reached levels comparable to those achieved in 10% fetal bovine serum cultures while maintaining serum-free conditions.
[Conclusion] This optimized system provides a reproducible and scalable platform for serum-free expansion of human T cells, supporting the manufacturing of ACT and chimeric antigen receptor T cell therapies.
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