Fig. 1: Induction of surface ectoderm and salivary gland organoids from hiPSCs.
a, Culture protocol of salivary gland differentiation from hiPSCs. TGF-β-i, inhibitor of transforming growth factor-β1. b, Real-time RT-PCR analysis of oral ectoderm genes in the aggregates on day 12 with or without BMP4 and SB431542 (SB). The data are normalized to ribosomal protein L27 and are presented as means ± s.d. (n = 3 biological replicates). Statistical significance was determined by one-way ANOVA followed by Tukey’s post-hoc test. c, Phase-contrast images of aggregates on days 2 (left) and 12 (right) without SB431542. Scale bars, 200 μm. The images are representative of three independent experiments. d, Phase-contrast images of aggregates on days 12 (left) and 20 (right) with SB431542 treatment. Scale bars, 200 μm. The images are representative of 18 independent experiments. e, Pan-CK and SOX9 were detected via immunofluorescence analysis. The images are representative of three samples showing similar results. Scale bars, 200 μm. f, Real-time RT-PCR analysis of SOX9 in the aggregates on days 12, 16 and 20. The data are normalized to ribosomal protein L27 and are presented as means ± s.d. (n = 4 biological replicates). Statistical significance was determined by one-way ANOVA followed by Tukey’s post-hoc test. g, Representative phase-contrast images of branching structures on day 60. Scale bars, 200 μm. h, Immunofluorescence for Pan-CK, SOX9 and AQP5 in the branching structures of day 60 aggregates. The images are representative of six independent experiments. Scale bars, 400 μm (top) and 50 μm (bottom).
