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. Author manuscript; available in PMC: 2024 Oct 23.
Published in final edited form as: Dev Cell. 2023 Aug 14;58(20):2163–2180.e9. doi: 10.1016/j.devcel.2023.07.013

Figure 5. Human Induced Pluripotent Stem Cells (HiPSC) Derived Pre-Ameloblast Differentiation Protocol Guided by sci-RNA-seq.

Figure 5.

(A) Schematic of the 16-day differentiation protocol targeting signaling pathways using growth factors and small molecules. (B) QRT-PCR analysis showing upregulated expression of oral epithelium markers PITX2 and KRT14 at Day 10 of differentiation. (C) Bulk RNA-seq analysis demonstrating upregulation of ameloblast markers SP6 and AMBN at Day 16 of differentiation compared to undifferentiated hiPSC control. (D) Evaluation of pathway efficiency during differentiation by removing each agonist and or adding FGFR-mb to inhibit the FGFR1/2c pathway and assessing AMBN expression in QRT-PCR. Each performed in duplicates or more. (E) Projection of Day 16 differentiated cells onto in vivo dental epithelium-derived cell types, showing that 60% of the cells resemble the gene expression pattern of PA and eAM. Error bars represent SEM. Statistical significance was determined using one-way ANOVA; ***p < 0.001; ****p < 0.0001.