成釉细胞微环境对胚胎干细胞特性的影响

目的研究胚胎干细胞诱导后成牙分化的可行性。方法使用成釉细胞无血清条件培养液诱导胚胎干细胞分化,观察诱导后细胞形态学变化以及体外检测基因表达和免疫表型分析。结果成釉细胞无血清条件培养液诱导胚胎干细胞后,拟胚体的外周有鹅卵石样的细胞,类似于上皮样的细胞生成。同时,诱导组细胞中可检测CK14、AMBN及AMGN mRNA的表达,DMP1和DSPP表达阴性,进一步免疫表型分析显示,诱导组胚胎干细胞表达CK14、AMBN及AMGN阳性。结论利用成釉细胞无血清条件培养液分泌的信号分子在体外模拟上皮的微环境,使得胚胎干细胞实现了向牙源性上皮细胞方向的表型转化。

Effect of ameloblast micro-environment on embryonic stem cell characteristics

Objective To study the feasibility of tooth differentiation induced by embryonic stem cells. Methods Embryonic stem cell differentiation was induced by ameloblast serum-free conditioned medium(ASF-CM), morphological changes were detected, gene expression and immunophenotype were detected in vitro. Results After being treated with ASF-CM, cobblestone-like cells, which resembled dental epithelial cells, were observed at the periphery of the plated mEBs. At the same time, in the cells of the induction group, CK14, AMBN and AMGN mRNA expression could be detected, and the expression of DMP1 and DSPP was negative. Further analysis showed that the expression of CK14, AMBN and AMGN in the immune phenotype of embryonic stem cells in the induction group was positive. Conclusion The embryonic stem cells are transformed into odontogenic epithelial cells by using the signal molecules secreted by ameloblast serum-free conditioned medium in vitro to simulate the epithelial microenvironment.