颅神经嵴干细胞成牙本质样细胞分化诱导的体内实验研究  

Odontoblast-like Cell Phenotype Differentiation of Cranial Neural Crest Stem Cell in vivo

作　　者：江宏兵[1] 田卫东[1] 刘磊[1] 汤炜[1] 郑晓辉[1] 陈希哲[1] 

机构地区：[1]四川大学华西口腔医院口腔疾病研究国家重点实验室

出　　处：《四川大学学报（医学版）》2008年第2期276-278,282,共4页Journal of Sichuan University(Medical Sciences)

基　　金：国家自然科学基金(批准号30471904);中国博士后科学基金(20060391019);教育部重点研究项目(106134)资助

摘　　要：目的观察诱导颅神经嵴干细胞(cranial neural crest stem cell,CNCSC)后的裸鼠体内成牙本质样细胞表型分化,探讨CNCSC用于牙再生研究的可行性。方法颅神经管组织块无血清条件培养法获取CNCSC,体外经成纤维生长因子8(FGF8)、骨形态发生蛋白2(BMP2)、转化生长因子β1(TGFβ1)、牙本质基质非胶原蛋白等因子诱导后,与胶原-壳聚糖三维凝胶支架复合植入裸鼠背部皮下,免疫组织化学方法检测型胶原表达、牙本质涎磷蛋白(DSPP)表达,鉴定其成牙本质样细胞表型分化特征。结果诱导后CNCSC植入体内后1月表现为型胶原及DSPP阳性表达,2月后DSPP表达增强,局部区域可见极性栅栏状排列的细胞分布,并可见明显的细胞外基质形成。结论CNCSC在体内可向成牙本质样细胞表型分化,为进一步开展牙再生研究奠定了基础。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, TGFβ1 and dentin matrix noncollagen 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.

关 键 词：颅神经嵴 成牙本质细胞 支架 牙再生 

分 类 号：R78[医药卫生—口腔医学]