Figure 4.

IPSC-NCSCs Are Not Immunosuppressive

(A) Schematic outline of transwell assay format. (B) Representative flow cytometry histograms show T cell proliferation, detected by quantifying levels of CFSE by flow cytometry. Anti-CD3/anti-CD28-stimulated PBMCs were co-cultured with BM-MSCs, NIBSC8-MSCs, and NIBSC8-NCSCs, and reduction of T cell proliferation was measured by quantifying CFSE^low cells (shown in pink) in total CD3⁺ T cells and CD3⁺CD8⁺ T cells. (C) Graphs show relative total CD3⁺ T cell and CD3⁺CD8⁺ T cell proliferation in a transwell assay format (tw) and without transwell inserts (w/o tw). Bars represent proliferation in PBMCs only, in stimulated (stim) PBMCs only, and in stim PBMCs in response to co-culture with BM-MSCs, NIBSC8-MSCs, and NIBSC8-NCSCs relative to stim PBMCs. Error bars represent ± SEM (w/o tw: n = 9 biological replicates in three separate experiments; with tw: n = 3 biological replicates in a single experiment). Ordinary one-way ANOVA with Bonferroni post-test was performed. Asterisks denote statistical significance in comparison with stim PBMCs: *p ≤ 0.05, **p ≤ 0.01, and ***p ≤ 0.001.