Abstract：
Primary cultured odontoblasts rapidly lose their tissue-specific phenotype. To identify transcription factors (TF) that are important for the maintenance of the odontoblast phenotype, primary cultures of C57BL/6 J mouse dental mesenchymal cells (DMC) were isolated, and expression of TF and odontoblast marker genes in cells immediately after isolation and 2 days after culture were comprehensively evaluated and compared using RNA-sequencing (RNA-seq). The expression of odontoblast markers in mouse dental mesenchymal cells decreased rapidly after isolation. In addition, the expression of Hedgehog-related, Notch-related, and immediate- early gene (IEG)-related transcription factors significantly decreased. Forced expression of these genes in lentiviral vectors, together with fibroblast growth factor 4 (FGF4), fibroblast growth factor 9 (FGF9), and the Wnt pathway activator CHIR99021, significantly induced the expression of odontogenic marker genes. These results indicate, for the first time, that Notch signaling and early genes may be important for maintaining odontoblast cultures. Furthermore, simultaneous stimulation of FGF, Wnt, Hedgehog, Notch pathways, and IEG transcription factors cooperatively promoted the maintenance of the odontoblast phenotype. These results suggest that the Hedgehog and Notch signaling pathways may play an important role in maintaining odontoblast phenotypes, in addition to FGF and Wnt signaling.
摘要：
原代培养的成牙本质细胞迅速失去其组织特异性表型。为了鉴定对成牙本质细胞表型的维持重要的转录因子(TF),分离C57BL/6J小鼠牙间充质细胞（DMC）的原代培养物，分离后立即和培养后2天，使用RNA测序（RNA-seq）全面评估和比较了TF和成牙本质细胞标记基因的表达。分离后，成牙本质细胞标志物在小鼠牙间充质细胞中的表达迅速下降。此外,刺猬相关的表达，缺口相关,早期基因(IEG)相关转录因子显著降低。这些基因在慢病毒载体中的强制表达，与成纤维细胞生长因子4（FGF4），成纤维细胞生长因子9(FGF9),和Wnt通路激活剂CHIR99021，显著诱导牙源性标记基因的表达。这些结果表明,第一次,Notch信号和早期基因可能对维持成牙本质细胞培养很重要。此外，同时刺激FGF，Wnt,刺猬,缺口通道,和IEG转录因子协同促进成牙本质细胞表型的维持。这些结果表明，Hedgehog和Notch信号通路可能在维持成牙本质细胞表型中起重要作用。除了FGF和Wnt信号。
