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. 2023 Jun 29;11:1164811. doi: 10.3389/fcell.2023.1164811

FIGURE 2.

FIGURE 2

Characterization of hiPSC-derived ameloblast organoids. (A) H&E staining of paraffin sections of hiPSC-derived organoids. The round organoids displayed dense and complex morphologies. Organoids consisted of one cell layer in the outermost layer, a dense inner structure, and budding structures that are irregularly entangled inward. (B, C) Immunostaining of epithelial cell markers, E-CAD and CK14, (D) a basal epithelial cell marker, p63, (E) a basement membrane marker, Col IV, (F) a dental epithelial cell marker, PITX2, and (G, H) ameloblast markers, DLX3 and AMELX in organoids. (B′–H′) Higher magnification of the regions of interest shown in (B–H). (I–M) mRNA expression of CK14, PITX2, SHH, DLX3, and AMELX was analyzed by RT-qPCR. On comparing the 2D (hiPSC-derived dental epithelial cells) and 3D cultures (hiPSC-derived organoids), the RNA expression levels of epithelial cell markers, dental epithelial cell markers, and ameloblast markers were significantly increased in the 3D culture. All experiments were performed in triplicate. Data are presented as means ± standard deviations (SD). TOPRO-3 (blue) was used to label the nuclei. Scale bar = 100 μm. White arrow heads: expression cells.