In vitro odontoblast-like cell differentiation of cranial neural crest cells induced by fibroblast growth factor 8 and dentin non-collagen proteins

During tooth development, cranial neural crest (CNC) cells represent a population of pluripotent stem cells that give rise to various dental tissues. This study aimed to investigate whether CNC cells could differentiate into odontoblast-like cells by in vitro induction. CNC cells were isolated from explants of cranial neural tubes and cultured in serum-free Dulbecco's modified Eagle's medium (DMEM)/F12 medium which contained fibroblast growth factor 8 (FGF8) and dentin non-collagen proteins (DNCP). The initiation of controlled differentiation was determined using histological assays, and the expression of specific gene phenotypes was detected using immunocytochemical staining and reverse transcription-polymerase chain reaction (RT-PCR). The first branchial arch phenotype of the CNC cells demonstrated negative Hoxa2 expression and positive vimentin expression in the presence of 100 ng/ml FGF8. Following DNCP induction, the CNC cells became bipolar, demonstrated high alkaline phosphatase (ALP) activity, and formed mineralized nodules. In addition, the expression of DSPP, DMP1, and collagen type I confirmed the odontoblast phenotype. The results indicate that the tissue-specific cellular differentiation (odontoblast-like cells) of early-stage embryonic-derived cells (such as CNC cells) can be induced by adult extracellular matrix proteins (such as DNCP). CNC cells may be used as a valuable cell model for research on dental tissue differentiation and regeneration.