Stage-specific role of Amelx activation in stepwise ameloblast induction from mouse induced pluripotent stem cells

Xinchao Miao, Kunimichi Niibe, Maolin Zhang, Zeni Liu, Praphawi Nattasit, Yumi Ohori-Morita, Takashi Nakamura, Xinquan Jiang, Hiroshi Egusa

Research output: Contribution to journalArticlepeer-review

Abstract

Amelogenin comprises ~90% of enamel proteins; however, the involvement of Amelx transcriptional activation in regulating ameloblast differentiation from induced pluripotent stem cells (iPSCs) remains unknown. In this study, we generated doxycycline-inducible Amelx-expressing mouse iPSCs (Amelx-iPSCs). We then established a three-stage ameloblast induction strategy from Amelx-iPSCs, including induction of surface ectoderm (stage 1), dental epithelial cells (DECs; stage 2), and ameloblast lineage (stage 3) in sequence, by manipulating several signaling molecules. We found that adjunctive use of lithium chloride (LiCl) in addition to bone morphogenetic protein 4 and retinoic acid promoted concentration-dependent differentiation of DECs. The resulting cells had a cobblestone appearance and keratin14 positivity. Attenuation of LiCl at stage 3 together with transforming growth factor β1 and epidermal growth factor resulted in an ameloblast lineage with elongated cell morphology, positivity for ameloblast markers, and calcium deposition. Although stage-specific activation of Amelx did not produce noticeable phenotypic changes in ameloblast differentiation, Amelx activation at stage 3 significantly enhanced cell adhesion as well as decreased proliferation and migration. These results suggest that the combination of inducible Amelx transcription and stage-specific ameloblast induction for iPSCs represents a powerful tool to highlight underlying mechanisms in ameloblast differentiation and function in association with Amelx expression.

Original languageEnglish
Article number7195
JournalInternational journal of molecular sciences
Volume22
Issue number13
DOIs
Publication statusPublished - 2021 Jul 1

Keywords

  • Ameloblast
  • Amelogenin
  • Cell adhesion
  • Cell differentiation
  • Induced pluripotent stem cells
  • Transcriptional activation

ASJC Scopus subject areas

  • Catalysis
  • Molecular Biology
  • Spectroscopy
  • Computer Science Applications
  • Physical and Theoretical Chemistry
  • Organic Chemistry
  • Inorganic Chemistry

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