Abstract：
Mesenchymal stem cells in the dental tissue indicate a disposition for differentiation into diverse dental lineages and contain enormous potential as the important means for regenerative medicine in dentistry. Among various dental tissues, the dental pulp contains stem cells, progenitor cells and odontoblasts for maintaining dentin homeostasis. The conventional culture of stem cells holds a limit as the living tissue constitutes the three-dimensional (3D) structure. Recent development in the organoid cultures have successfully recapitulated 3D structure and advanced to the assembling of different types. In the current study, the protocol for 3D explant culture of the human dental pulp tissue has been established by adopting the organoid culture. After isolating dental pulp from human tooth, the intact tissue was placed between two layers for Matrigel with addition of the culture medium. The reticular outgrowth of pre-odontoblast layer continued for a month and the random accumulation of dentin was observed near the end. Electron microscopy showed the cellular organization and in situ development of dentin, and immunohistochemistry exhibited the expression of odontoblast and stem cell markers in the outgrowth area. Three-dimensional explant culture of human dental pulp will provide a novel platform for understanding stem cell biology inside the tooth and developing the regenerative medicine.
摘要：
牙齿组织中的间充质干细胞表明分化成多种牙齿谱系的倾向，并且作为牙科再生医学的重要手段具有巨大的潜力。在各种牙齿组织中，牙髓含有干细胞，维持牙本质稳态的祖细胞和成牙本质细胞。干细胞的常规培养存在限制,因为活组织构成三维(3D)结构。类器官培养的最新发展已成功地概括了3D结构，并发展到不同类型的组装。在目前的研究中,通过采用类器官培养，建立了人牙髓组织3D外植体培养的方案。从人类牙齿中分离出牙髓后，在添加培养基的情况下,将完整组织置于用于Matrigel的两层之间。成牙本质层的网状生长持续了一个月，并且在接近尾声时观察到牙本质的随机积累。电子显微镜显示牙本质的细胞组织和原位发育，免疫组化显示成牙本质细胞和干细胞标志物在生长区的表达。人类牙髓的三维外植体培养将为理解牙内干细胞生物学和发展再生医学提供一个新的平台。
