Department of Stomatology, Xinqiao Hospital, Third Military Medical University, Chongqing 400037, PR China.
Arch Oral Biol. 2009 Mar;54(3):216-22. doi: 10.1016/j.archoralbio.2008.10.003. Epub 2008 Dec 6.
Notch signalling controls cell fate decisions in adult and embryonic tissues. The Notch ligand Delta1 is known to influence proliferation and differentiation of many kinds of tissue specific stem cells. In the present study, we investigated the role of Delta1 in the regulation of dental pulp stem cells (DPSCs) in vitro.
DPSCs were isolated from impacted third molars. Expression of human Notch1, 2 and Delta1 in DPSCs were detected by immunochemistry. Delta1 overexpressed DPSCs were constructed by a retroviral method. Delta1 transduced DPSCs proliferation changes were examined by means of colony-forming assay, BrdU incorporation assay and cell cycle analysis. Delta1 transduced DPSCs were cultured in differentiation-inductive medium. The nodule formation and DSPP expression were evaluated.
It was shown that the Notch receptors and Delta1 ligand were expressed throughout the proliferation and differentiation process of cultured dental pulp stem cells. Furthermore, it was found in our study that Delta1 could significantly enhance the proliferation of DPSCs and permit DPSCs differentiating into odontoblast-like cells in differentiation-inductive environments.
Our findings verified that Notch-Delta1 signalling was expressed in human DPSCs in vitro and appeared to play pivotal role in DPSCs proliferation enhancement and differentiation regulation, thereby consistent with the hypothesis that the Notch pathway controls stem cell fate during pulp regeneration.
Notch 信号通路在成人和胚胎组织中控制着细胞命运决定。已知 Notch 配体 Delta1 影响许多组织特异性干细胞的增殖和分化。本研究旨在探讨 Delta1 在体外调节牙髓干细胞(DPSCs)中的作用。
从阻生第三磨牙中分离 DPSCs。通过免疫化学方法检测 DPSCs 中人 Notch1、2 和 Delta1 的表达。通过逆转录病毒方法构建 Delta1 过表达 DPSCs。通过集落形成试验、BrdU 掺入试验和细胞周期分析检测 Delta1 转导 DPSCs 增殖变化。将 Delta1 转导的 DPSCs 培养在诱导分化培养基中,评估结节形成和 DSPP 表达。
结果表明,Notch 受体和 Delta1 配体在培养牙髓干细胞的增殖和分化过程中均有表达。此外,本研究还发现 Delta1 可显著增强 DPSCs 的增殖,并允许 DPSCs 在诱导分化环境中分化为成牙本质细胞样细胞。
我们的研究结果证实,Notch-Delta1 信号通路在人 DPSCs 中体外表达,并在 DPSCs 增殖增强和分化调节中发挥关键作用,这与 Notch 通路在牙髓再生过程中控制干细胞命运的假说一致。