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目的:研究成纤维细胞生长因子8(FGF8)对成人牙髓干细胞(hDPSCs)定向分化为成牙本质细胞及牙髓组织的影响。方法:首先分离、克隆培养hDPSCs,通过流式细胞术检测细胞表面标志物鉴定hDPSCs;矿化液中添加50μg/L的FGF8诱导hDPSCs分化,通过real-time PCR检测分化后的细胞中牙本质涎磷蛋白(DSPP)、碱性磷酸酶(ALP)、骨涎蛋白(BSP)和核心结合因子α1(Cbfa-1)在mRNA水平的表达;E11.5小鼠牙源性上皮联合FGF8与hDPSCs细胞团重组,再将组织块种植于裸鼠肾囊膜下培养,通过DNA原位杂交鉴定成牙本质细胞及牙髓细胞的来源。结果:成功分离培养hDPSCs,其表面标志物CD29和CD90呈阳性表达;经FGF8诱导的hDPSCs形成较明显的矿化结节,并且牙本质特异性蛋白DSPP、BSP及Cbfa-1表达量上调;E11.5小鼠牙源性上皮联合FGF8可以诱导hDPSCs分化为成牙本质细胞及牙髓细胞。结论:FGF8能够辅助牙源性上皮定向诱导hDPSCs分化为成牙本质细胞及牙髓细胞,并形成牙本质及牙髓腔结构。
Objective: To investigate the effect of fibroblast growth factor 8 (FGF8) on the differentiation of adult dental pulp stem cells (hDPSCs) into odontoblasts and dental pulp. Methods: hDPSCs were isolated, cloned and cultured, and cell surface markers were detected by flow cytometry to identify hDPSCs. FHGs differentiated into hDPSCs by adding 50μg / L of FGF8 in mineralized solution, and the dentin salivary glands in differentiated cells were detected by real-time PCR (DSPP), alkaline phosphatase (ALP), bone sialoprotein (BSP) and corebinding factor α1 (Cbfa-1) at the mRNA level; E11.5 mouse dental epithelium combined with FGF8 and hDPSCs cells After reorganization, the tissue was planted in the subrenal capsule of nude mice. The origin of odontoblasts and dental pulp cells was identified by DNA in situ hybridization. Results: The hDPSCs were successfully isolated and cultured, and the surface markers CD29 and CD90 were positive. The hGMPCs induced by FGF8 formed more obvious mineralized nodules and the expression of dentin-specific proteins DSPP, BSP and Cbfa-1 were up-regulated. E11 .5 Mice odontogenic epithelium combined with FGF8 can induce hDPSCs to differentiate into odontoblasts and dental pulp cells. Conclusion: FGF8 can assist the differentiation of hDPSCs into odontoblasts and odontoblasts and induce the dentin and pulp cavity structure.