hiPSC-derived Neurosphere Transplantation

The YZWJs513 hiPSC line was used throughout this study. NS/PCs were prepared at a good manufacturing practice-grade cell processing facility at Osaka National Hospital, Japan. The clinical-grade human leukocyte antigen super-donor-derived, integration-free hiPSC line, YZWJs513, established by the iPSC Stock Project and organized by the Kyoto University Center for iPS Cell Research and Application, was used. The cells were cultured for 4 days in a floating culture, and the neurospheres were used for transplantation (Fig. 1A). Cells were treated with small-molecule GSI and N-[N-(3,5-difluorophenacetyl)-l-ananyl]-S-phenylglycine t-butyl ester (DAPT) (10 μM, Sigma-Aldrich, Inc., D5942) for 1 day before transplantation, as described in a previous study^{13 (link)}. Detailed methods are provided in the Supplementary Material.