Fig. 5: Functional recovery of FVIII in cell-transplanted HA mice.
a Reverse transcription PCR (RT‒PCR) analysis was used to detect the expression of human FVIII in nontransplanted and transplanted HA mice after intravenous transplantation. Human ACTIN was used to identify transplanted ECs. Mouse Gapdh was used as a control. b FVIII activity was measured in blood plasma harvested from nontransplanted and transplanted HA mice after intravenous transplantation. The data represent the activity detected per 1 × 106 ECs. HA (nontransplanted HA mice, n = 3 for each day), Patient (n = 3 for each day), and Corrected (n = 6, 3, 4, 3, and 3). The data are the means ± SEMs. *p < 0.05 and ***p < 0.001 compared with mice with ECs differentiated from parental iPSCs at each timepoint (two-way ANOVA with post hoc Dunnett’s test). c Survival curves of nontransplanted and intravenously transplanted HA mice after the tail clip assay. HA (n = 6), Patient (n = 6), Corrected (n = 18). *p < 0.001 compared with the patient group (log-rank test). d The survival of subcutaneously transplanted cells in immunodeficient mice was confirmed. ECs differentiated from luciferase reporter KI iPSCs were transplanted subcutaneously into the thighs of immunodeficient mice, and cell survival was monitored by weekly in vivo live imaging up to 13 weeks after transplantation (left panel, n = 4). Luciferase activity was still detected 13 weeks after transplantation (right panel). e The long-term therapeutic efficacy of subcutaneously transplanted ECs differentiated from parental or corrected iPSCs was determined at 4 and 13 weeks after subcutaneous transplantation into an immunodeficient hemophilia model. Patient (n = 3), Corrected (n = 4). *p < 0.05, **p < 0.01 compared with the patient group (Student’s t test).
