Fig. 3: T-cell regeneration from tumor-specific TI-CTL-derived iPSC.

a Overview of the TIL-iPSC establishment and T-cell regeneration. TIL-iPSC were established from tumor-specific TI-CTL. Sendai virus was used for the reprogramming. Feeder-free protocols were applied for the T-cell regeneration from TIL-iPSC. HPC hematopoietic progenitor cell. b Flow cytometry analysis regarding T-cell lineage markers at each differentiation stage. Left panels were gated on CD45⁺ live cells. Right panels were gated on CD45⁺CD4⁻CD8α⁺ live cells. c Phenotype-related markers on TIL-iPS-T. Representative data of four independent experiments. Data were acquired from C-T-11 Vβ5.1 clone. All data were gated on CD45⁺CD4⁻CD8α⁺CD8β⁺ live cells. d TCF-1 (TCF-7) expression on TIL-iPS-T. TI-CTL and TIL-iPS-T data were acquired from C-T-11 Vβ5.1 clone. PB-CTL (peripheral blood-derived CTL) were from a healthy donor. Left: quantification by qPCR. mRNA was extracted from the CD45⁺CD4⁻CD8α⁺CD8β⁺ live cell population. Representative data of two independent experiments are shown. The expression levels were standardized with the ΔCT method by ACTB expression and further standardized with the ΔΔCT method by the ΔCT mean of TCF-1 expression on TI-CTL. ****P < 0.0001, two-tailed unpaired t test. Dots represent individual data. Means are shown; error bars represent SD. n = 3 per sample. Right: Representative flow cytometry data of two independent experiments are shown. Data were gated on CD45⁺CD4⁻CD8α⁺CD8β⁺ cells. MFI mean fluorescence intensity. e Mitochondria biomass on TIL-iPS-T. Data were acquired from C-T-11 Vβ5.1 clone. Histograms were gated on CD45⁺CD4⁻CD8α⁺CD8β⁺ live cells. Representative data of four independent experiments are shown.