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Updated: Jul 3, 2026

Cranial Neural Crest Cells Three-Dimensional In Vitro Differentiation Protocol for Multiplexed Assay
Published on: February 14, 2025
[Odontoblast-like cell phenotype differentiation of cranial neural crest stem cell in vivo]
Hong-Bing Jiang1, Wei-Dong Tian, Lei Liu
1State Key Laboratory of Oral Diseases, West China Stomatological Hospital, Sichuan University, Chengdu 610041, China.
Objective:
To investigate the feasibility of tooth regeneration by seeding cranial neural crest stem cell (CNCSC) in vivo.
Methods:
Cranial neural tubes, dissected from mouse E9 d, were explanted onto fibronectin-coated dishes. CNCSC emigrated from the explanted neural tubes, and were cultured in a free-serum medium containing modified DMEM/F12. CNCSC, induced by FGF8, BMP2, TGFbeta1 and dentin matrix non-collagen protein (DMNCP), were cultured with collagen/chitosan, and implanted into the subcutaneous part of immunodeficiency mouse. The expression of collagen I/dentin sialophosphoprotein (DSPP) was analyzed by immunocytochemistry.
Results:
With the scaffolds destroying, columnar cells possessing polarized nuclei and matrix produced by cells were showed in some regions. Immunohistochemical staining demonstrated that collagen type I and DSPP were expressed throughout the cytoplasm and matrix produced by cells.
Conclusion:
By tissue engineering approach, our experiments further verify the odontoblast-like cell phenotype differentiation of CNCSC in vivo.
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