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人表皮干细胞可以作为牙齿再生中上皮源性的种子细胞,但是其成釉分化的效率低下.本研究分离培养了人牙胚上皮细胞,利用E13.5的小鼠牙间充质与其重组,构建重组牙胚,对其成釉分化的潜能和机制进行研究.研究结果发现,体外培养的P1代人牙胚上皮细胞成釉率高达50%.随着传代次数的增加,成釉率明显下降.通过对牙上皮发育分化相关基因的表达检测和分析表明,重组牙胚成牙分化能力和成釉潜能的下降与牙上皮发育相关基因的表达状态密切相关.特别是FGF8表达水平的下调以及PITX2不同亚型在人牙胚细胞中表达量的不均衡,可能是导致人牙胚细胞成釉潜能下降并丧失的主要原因.本研究结果为理解牙齿再生过程中上皮源性的种子细胞的成釉机制提供了新的实验数据,对进一步提高表皮干细胞在牙齿再生过程中的成釉率有指导意义.
Human epidermal stem cells can be used as epithelial-derived seed cells in tooth regeneration, but their efficiency of glaze differentiation is low.In this study, human tooth germ epithelial cells were isolated and cultured, The results showed that the rate of glaze formation of P1 tooth germ epithelial cells in vitro was as high as 50%, and the rate of glaze formation decreased significantly with the increase of passage times. Detection and analysis of the expression of genes related to the development and differentiation of the dental epithelium showed that there is a close relationship between the expression of tooth epithelial differentiation and the odontogenic potential and the potential of ameloblasts, especially the down-regulation of FGF8 expression and PITX2 The imbalanced expression of subtypes in human tooth germ cells may be the main reason leading to the loss of enamel potential and loss of human tooth germ cells.This study is to understand the mechanism of enamel formation of epithelial-derived seed cells during tooth regeneration Provides new experimental data, and is of guiding significance for further improving the rate of glaze formation during the tooth regeneration process of epidermal stem cells.