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Fig. 4 | Stem Cell Research & Therapy

Fig. 4

From: Generation of individualized immunocompatible endothelial cells from HLA-I-matched human pluripotent stem cells

Fig. 4

In vitro T cell stimulations by HLA-I-modified iPSC-ECs. A Schematic diagram of the protocol for T cell proliferation induced by co-culture of PBMCs with the HLA-I-modified allogeneic iPSC-ECs. B HLA genotyping of C55 iPSCs and donor PBMCs. Table shows HLA-I genotyping (A, B and C alleles). C T cell proliferation assay with FCM: the percentage of proliferating T cells were plotted by gating of the reduced CFSE fluorescence. CD3+ (upper panel), and CD8+ (lower panel) T cell populations co-cultured with different iPSC-ECs (WT, KO, or A11); the unmodified iPSC-ECs from C55 were called as WT group, B2M KO ones from C55 as KO group and HLA-A11-modified ones from C55-B2MKO as A11 group. T cells cultured alone were used as negative controls (Neg.Ctrl.); T cells activated with CD3 and CD28 antibodies served as positive controls (Pos.Ctrl.). D Statistical data for T cell proliferation: scatterplots displaying percentage of proliferating T cells in CD3+ (left, n = 3 independent experiments) and CD8+ (right, n = 3 independent experiments) T cell populations when co-cultured with WT, KO, or A11 iPSC-ECs. Control groups as described in C. Statistical significance was determined with paired one-way ANOVA followed by Tukey’s multiple comparison test. Data are means ± SEM; ***p < 0.001, **p < 0.01

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