Additional Supporting Information may be found in the online version of this article.

STEM_1415_sm_SuppFigure1.pdf433KSupplemental Figure 1. Characterization of hiPSCderived neural subpopulations. Panel A. FOXA2/TH/Cb and FOXA2/TH/serotonin- expression in unsorted, NCAM+, NCAM+/CD29low and NCAMlow-population. B. Cell counts for FOXA2/TH, TH/Cb/FOXA2, Cb, and serotonin-positive cells in unsorted, NCAM+, NCAM+/CD29low, and NCAMlow population. Significant differences between unsorted and sorted cell populations are presented, * p<0.05. Statistical analysis was performed with Kruskall-Wallis test (non-parametric ANOVA) and Dunn's multiple comparison test between selected pairs. Panel C. FOXA2/TH, EN1/TH, β-tubulinIII, transthyretin/NCAM and SSEA-4/Tra 1-60 expression in unsorted and NCAM+/CD29low populations. Scale bar 50 μm.
STEM_1415_sm_SuppFigure2.pdf597KSupplemental Figure 2. In vivo survival of unsorted and NCAM+/CD29low PiPSCderived neural cell grafts. Panel A. Representative images of NCAM- positive unsorted PiPSC-derived neural cell grafts in 6-OHDA rats striatum 16 weeks after transplantation (n=5). Panel B. Representative images of NCAM-positive NCAM+/CD29low PiPSC-derived neural cell grafts in 6-OHDA rats striatum 16 weeks after transplantation (n=5).
STEM_1415_sm_SuppFigure3.pdf435KSupplemental Figure 3. Density analysis of THpixel intensity in 6OHDA lesioned rats striatum 16 weeks after PiPSCderived DA neuron transplantation. TH expression in; A. 6-OHDA lesion control rat, B. rat striatum transplanted with unsorted PiPSC-derived DA neurons, C. rat striatum transplanted with NCAM+/CD29low PiPSC-derived DA neurons, D. intact striatum (positive control for TH-staining). E. ImageJ analysis of TH-pixel density in 6-OHDA lesioned striatum. Analyzed animal groups consisted of: lesion-control without graft (n=8), unsorted cell grafts (n=6) or NCAM+/CD29low cell grafts (n=8) in striatum, and intact striatum (n=9). Statistically significant differences in the striatal TH+ pixel density were detected between lesion control animals and lesioned animals with DA neuron cell grafts, * p<0.05. Scale bar in panels A-D = 600μm.
STEM_1415_sm_SuppFigure4.pdf588KSupplemental Figure 4. Survival and functionality of PiPSCderived DA neurons in 6OHDAlesioned rat striatum. A. PiPSC–derived DA neurons differentiated with modified Method C (see materials and methods), expressed FOXA2/TH/hNCAM 6 weeks after transplantation (lines MF27.04 and MF66.02). B. 20 weeks after transplantation FOXA2/TH/NCAM and FOXA2/TH/Girk2-positive cells were detected in the cell graft (line MF27.04). C. 20 weeks after transplantation PiPSC–derived neural grafts were negative for transthyretin. Transplanted cells were detected with human nuclear marker (red) in the rat striatum. D. Amphetamine-induced rotation test showed that 6-OHDA rats transplanted with PiPSC-derived DA- neurons had decreased number of rotations related to baseline and control group 4 months after transplantation (* p<0.05, lines MF27.04 and MF66.02 n=6/line, 6-OHDA lesioned rats without graft were used as a control group n=8). Scale bar 100μm A and C, scale bar 50μm B.
STEM_1415_sm_SuppTable1.pdf59KSupporting Information
STEM_1415_sm_SuppTable2.pdf67KSupporting Information
STEM_1415_sm_SuppTable3.pdf55KSupporting Information
STEM_1415_sm_SuppTable4.pdf65KSupporting Information

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