Ameloblast Differentiation In Vitro: A Step Closer to Enamel Engineering

体外成釉细胞分化：离牙釉质工程又近了一步

• 批准号: 7304940
• 负责人: Pamela K Den Besten
• 金额: $ 19.26万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-08-02 至 2009-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7304940
• 关键词: Ameloblastoma; Ameloblasts; Biology; Biomedical Engineering; Calcium; Cell Communication; Cell Differentiation process; Cell Lineage; Cell Maintenance; Cells; Condition; Cytokeratin-14 Staining Method; Dental; Dental Enamel; Differentiation and Growth; Enamel Formation; Enamel Organ; Engineering; Epithelial; Epithelial Cells; Epithelium; Extracellular Matrix; Extracellular Matrix Proteins; Goals; Growth; Growth Factor; Health; Human; Human body; In Vitro; Inner Enamel Epithelium; Knowledge; Lead; Life; MMP-20; Mesenchymal; Modeling; Natural regeneration; Organ; Pathology; Phenotype; Proteins; Reporter; Research; Resistance; Staging; System; Time; Tissues; Tooth structure; Undifferentiated; Up-Regulation; amelogenin; cellular engineering; dentin sialoprotein; fluorosis; improved; kallikrein 4; mineralization

DESCRIPTION (provided by applicant): Enamel is a mineralized tissue formed by the epithelially derived ameloblasts. The reciprocal interactions between epithelial cells of the enamel organ and dental mesenchymal cells lead to the differentiation of ameloblasts, which govern the synthesis of the enamel matrix. Studies of ameloblast function and differentiation have been limited by difficulties in isolation and maintenance of the cells in primary culture, as well as our limited knowledge of ameloblast biology. The goal of these proposed studies is to characterize ameloblast lineage cells in culture and to study their differentiation. We hypothesize that factors present in the local microenvironment of the ameloblasts will direct their differentiation and propose the following specific aims. Specific Aim1: To isolate, expand and maintain primary undifferentiated ameloblast lineage cells from single-cell colonies in vitro. Specific aim 2: To induce differentiation of ameloblast lineage cells to mature ameloblasts. Cells from the enamel organ epithelium will be cloned from single cells, characterized by the epithelial marker cytokeratin 14, and markers of preameloblasts including dentin sialoprotein (DSP) and amelogenin. Ameloblast differentiation will be induced by cell/cell interactions with dental mesencymal cells, extracellular matrix proteins, growth factors and calcium. The early-differentiated secretory ameloblasts will be characterized by upregulation of amelogenin, ameloblastin, and MMP-20. Mature ameloblasts will be characterized by upregulation of kallikrein 4 (KLK4), amelotin and Ae2. The identification of factors that drive ameloblast differentiation will allow us to better understand normal and pathological ameloblast biology, and how these cells engineer the formation of the enamel matrix. The regeneration of tooth enamel requires an understanding of the factors that control expansion and differentiation of ameloblast (enamel forming) lineage cells. In this proposal we propose to identify key factors that direct ameloblast lineage cell fate decisions as they form an enamel matrix.

描述（由申请人提供）：牙釉质是由上皮来源的成釉细胞形成的矿化组织。牙釉质器官的上皮细胞和牙齿间充质细胞之间的相互作用导致成釉细胞的分化，从而控制牙釉质基质的合成。由于原代培养细胞的分离和维持困难以及我们对成釉细胞生物学知识的有限，对成釉细胞功能和分化的研究受到限制。这些拟议研究的目标是表征培养物中的成釉细胞谱系细胞并研究它们的分化。我们假设成釉细胞局部微环境中存在的因素将指导它们的分化，并提出以下具体目标。具体目标1：在体外从单细胞集落中分离、扩增和维持原代未分化成釉细胞谱系细胞。具体目标2：诱导成釉细胞谱系细胞分化为成熟成釉细胞。釉质器官上皮细胞将从单细胞中克隆出来，其特征是上皮标记物细胞角蛋白 14 和前成釉细胞标记物，包括牙本质唾液酸蛋白 (DSP) 和釉原蛋白。成釉细胞分化将通过细胞/细胞与牙间质细胞、细胞外基质蛋白、生长因子和钙的相互作用来诱导。早期分化的分泌性成釉细胞的特征是釉原蛋白、成釉细胞蛋白和 MMP-20 的上调。成熟的成釉细胞的特点是激肽释放酶 4 (KLK4)、amelotin 和 Ae2 上调。识别驱动成釉细胞分化的因素将使我们能够更好地了解正常和病理性成釉细胞生物学，以及这些细胞如何设计牙釉质基质的形成。牙釉质的再生需要了解控制成釉细胞（牙釉质形成）谱系细胞扩张和分化的因素。在本提案中，我们建议确定在成釉细胞形成牙釉质基质时指导成釉细胞谱系细胞命运决定的关键因素。