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.
Jiang
Hong-Bing
HB
College of Stomatology, Nanjing Medical University, Nanjing, Jiangsu 210029, PR China. jhbtooth@yahoo.com.cn
Tian
Wei-Dong
WD
Liu
Lai-Kui
LK
Xu
Yan
Y
D000818:Animals; D002113:Calcification, Physiologic; D002454:Cell Differentiation; D016895:Culture Media, Serum-Free; D003804:Dentin / Q000378:metabolism; D016326:Extracellular Matrix Proteins / Q000494:pharmacology*; D051524:Fibroblast Growth Factor 8 / Q000494:pharmacology*; D051379:Mice; D008807:Mice, Inbred BALB C; D009432:Neural Crest / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D009804:Odontoblasts / Q000166:cytology*; D039904:Pluripotent Stem Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D012886:Skull / Q000166:cytology / Q000196:embryology*
D016895:Culture Media, Serum-Free; D016326:Extracellular Matrix Proteins; D051524:Fibroblast Growth Factor 8