Department of Endodontics, Tokyo Dental College, 2-9-18 Kanda-Misaki-Chou, Chiyoda, Tokyo, 101-0061, Japan.
Department of Biochemistry, Tokyo Dental College, 2-9-18 Kanda-Misaki-Chou, Chiyoda, Tokyo, 101-0061, Japan.
Med Mol Morphol. 2024 Dec;57(4):253-267. doi: 10.1007/s00795-024-00389-w. Epub 2024 Jul 11.
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.
原代培养的成牙本质细胞会迅速失去其组织特异性表型。为了鉴定对维持成牙本质细胞表型重要的转录因子（TF），我们分离了 C57BL/6J 小鼠牙间充质细胞（DMC）的原代培养物，并使用 RNA 测序（RNA-seq）全面评估和比较了细胞分离后立即和培养 2 天后 TF 和牙本质细胞标志物基因的表达。小鼠牙间充质细胞中成牙本质细胞标志物的表达在分离后迅速下降。此外，Hedgehog 相关、Notch 相关和即刻早期基因（IEG）相关转录因子的表达显著降低。这些基因在慢病毒载体中的强制表达，加上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，显著诱导了牙源性标志物基因的表达。这些结果首次表明，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，FGF、Wnt、Hedgehog、Notch 通路和 IEG 转录因子的同时刺激协同促进了成牙本质细胞表型的维持。这些结果表明，Hedgehog 和 Notch 信号通路可能除了 FGF 和 Wnt 信号外，在维持成牙本质细胞表型方面也起着重要作用。
