Fig. 3

Co-stimulatory blockade effectively prevents rejection to iPSC-grafts. (A) Schematic overview of CBA/N or C57BL/6 Luc-iPSC transplantation. (B) Bioluminescence images of C57BL/6 (upper panels) or CBA/N (lower panels) Luc-iPSC transplanted mice that received non-treatment, TDC + mAb, or CB + rapa. n = 3-5 per group. (C, D) Quantitative bioluminescence intensity of C57BL/6 (C) or CBA/N (D) Luc-iPSC-transplanted mice that received non-treatment, TDC + mAb, or CB + rapa. n = 3-5 per group. *p < 0.05, **p < 0.01 (Tukey’s HSD test). (E) Hematoxylin and eosin stained sections of C57BL/6 (left panel) or CBA/N (right panel) Luc-iPSC-derived teratomas under immunosuppression with TDC + mAb or CB + rapa on day 35. Lower panels show enlargement of insets in the upper panels. Scale bars: 100 μm. (F) Recipient T cell response in MHC-matched but minor antigen-mismatched iPSC transplantation. The T cell proliferation rates in each treatment group were normalized to that of C3129F1 mice stimulated with autologous irradiated splenocytes. Error bars indicate standard error of technical triplicates. Similar results were obtained in two independent experiments. *p < 0.05, **p < 0.01 (Tukey’s HSD test). (G) De novo anti-donor antibody production in the recipients. Error bars indicate standard error of biological replicates (n = 3, non-treatment; n = 5, TDC + mAb; n = 5, CB + rapa). Luc, luciferase; TDC, three drug combination; mAb, monoclonal antibody; CB, co-stimulatory molecule blocking; rapa, rapamycin; POD, post-operative day