Figure 3.

IPSC-NCSCs Fail to Stimulate Lymphocytes

(A) Representative histograms show levels of CD3⁺CD8⁺ T cell proliferation of responder cells after 5-day co-culture with autologous (auto), allogeneic (allo) PBMCs, NIBSC8-NCSCs, NIBSC35-NCSCs, or NIBSC8-SMCs as stimulator cells. (B) Graphs show stimulation index (SI) of total T cell (CD3⁺) and CD3⁺CD8⁺ T cell proliferation in the responder cell population in response to different stimulator cells. SI is calculated as the number of Ki67⁺ cells of each one-way MLR over the number of Ki67⁺ cells in an autologous control. Numbers are expressed relative to the control group: autologous (auto) PBMCs. A response was considered positive when the SI was ≥2. Error bars represent ± SEM (n = 11 biological replicates from three separate experiments). Ordinary one-way ANOVA with Bonferroni post-test was performed. Asterisks denote statistical significance: *p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001. (C) Supernatants of 5-day co-cultures were assayed for IFN-γ, IL-1β, IL-2, IL-6, IL-10, IL12-p70, and TNF-α using multiplex immunoassays. Error bars represent ± SEM (n = 10 biological replicates from three separate experiments). Ordinary one-way ANOVA with Bonferroni post-test was performed. Asterisks denote statistical significance: *p ≤ 0.05, **p ≤ 0.01, and ***p ≤ 0.001. Only statistically significant comparisons are shown in the graphs.