Fig. 3

Candidate T cells were screened and obtained utilizing modified culture conditions. A Phenotypic analysis of T cells was performed based on the expression of CD45RA, CCR7, CD122 and CD95 after exposure to either IL-2 or a combination of IL-7 and IL-15. The percentage of T_SCM (CD45RA⁺ CCR7⁺ CD122⁺ CD95⁺) cells was shown. Data represented as mean ± SD. N = 9 independent biological replicates. B T cells which were induced by either IL-2 or the combination of IL-7 and IL-15 were mixed with Karpas 299 target cells to conduct the cytotoxicity assay. Data represented as mean ± SD. N = 3 independent biological replicates. C Expansion of CD8⁺ T cells derived from 20 healthy donors within OptiVitro T-SFM medium containing IL-7 and IL-15. D and E The viability and expansion of five dominant CD8⁺ T cells, along with one suboptimal CD8⁺ T cells, following cryopreservation and thawing cycles. F and G TCR⁻/HLA-I⁻ CD8⁺ T cells derived from five dominant donors were transduced with CD5-CD30 CAR and incubated with Karpas 299 cells for 18 to 24 h. IFN-γ secretion within each group was analyzed utilizing ELISPOT assays. Data in (A) were analyzed by Student’s t-test. Data in (B) were analyzed by two-way ANOVA with Sidak’s multiple comparisons test. Data in (G) were analyzed by one-way ANOVA with Tukey’s multiple comparisons test. * p < 0.05, ** p < 0.01, *** p < 0.00