Fig. 1

General scheme of ESC differentiation and experimental outline of study. a After two passages on gelatin, ESC differentiation started with EB formation, followed by selection of Nestin⁺ cells in ITSFn medium and subsequent expansion in DMEM/F12 + B27 medium in the presence of bFGF. On day 24 of differentiation, cells were purified by FACS and MACS to yield a population of SSEA-1^–/PSA-NCAM⁺ progenitors, which were labeled with BrdU for 48 hours prior to transplantation. b Transplantation was performed on DPO7, the surgery for biotin dextran tracing took place on DPO21, and the von Frey filament test immediately before perfusion on DPO42. Motor behavior by BMS scoring was tested 1 day prior to SCI (DPO–1) and on DPO1, 6, 8, 10, 14, 21, 28, 35, and 42. bFGF basic fibroblast growth factor, BrdU 5-bromo-2′-deoxyuridine, DMEM Dulbecco’s modified Eagle’s medium, DPO days post operation, EB embryoid body, ESC embryonic stem cell, FACS, fluorescence-activated cell sorting, ITSFn insulin, transferrin, selenium chloride, fibronectin, MACS magnetic-activated cell sorting, PSA-NCAM polysialylated neural cell adhesion molecule, SCI spinal cord injury, SSEA-1 stage-specific embryonic antigen-1