Characterization of Corneal Epithelial Stem Cells

角膜上皮干细胞的表征

• 批准号: 6525116
• 负责人: Mary Ann Stepp
• 金额: $ 30.4万
• 依托单位: GEORGE WASHINGTON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-09-01 至 2006-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6525116
• 关键词: basement membrane; biomarker; cell adhesion; cell differentiation; cell proliferation; cellular pathology; corneal epithelium; flow cytometry; human tissue; integrins; laboratory mouse; laboratory rat; limbic system; microarray technology; peptide library; stem cells

DESCRIPTION (provided by applicant): The long term goal of this project is to use molecular approaches to define a set of cell specific surface narkers for human as well as rodent limbal stem cells and to use those markers as tools to characterize stem cells for the corneal epithelium in humans, rats, and mice in health and disease. Clinical experience and practice has shown us that limbal stem cells serve as the source for the cells comprising the corneal epithelium. At present, experimental confirmation that cells in the limbus are stem cells relies upon data from studies of vitro cell proliferation capacity and on the identification of a subset of these cells as slow-cycling. We hypothesize that a specific repertoire of cell surface markers exists which can define and select stem cells within the corneal limbus. Identification of such markers will aid in the study of the human ocular surface by providing tools to use to both select for and to target specific drugs or genes to these cells. Further, they will provide tools to allow for the analysis of limbal stem cell properties both in vivo and in vitro. The specific aims of this application are: 1) Identification of corneal stem-like cells from donor human corneas by selection based on cell adhesion properties, integrin expression profiles, and the ability of cells to proliferate in vitro, 2. Using phage displayed peptide libraries, identification of peptides that bind specifically to human, mouse, and rat limbal basal cells and use of those peptides to isolate and characterize stem cell specific surface markers, 3. In the mouse and rat, determine whether the slow-cycling cells observed in the limbus after neonatal BrdU administration possess similar cell surface markers as do human corneal stem cells, and 4. Using the knowledge and tools developed in Aims 1-3, characterize the limbal stem cells in the mouse under conditions where their differentiation and/or proliferation is altered in wild-type and in genetically engineered mice exhibiting symptoms of stem cell disease. Discovering specific molecular markers for the limbal stem cells will aid in the development of treatments for stem cell deficiency diseases of the ocular surface in humans.

描述（由申请人提供）：该项目的长期目标是 使用分子方法定义一组细胞特异性表面标记 人类以及啮齿动物的角膜缘干细胞，并使用这些标记作为工具 表征人类、大鼠和小鼠的角膜上皮干细胞 健康和疾病。临床经验和实践告诉我们，角膜缘 干细胞是构成角膜上皮的细胞的来源。 目前实验证实角膜缘细胞为干细胞 依赖于体外细胞增殖能力研究的数据以及 将这些细胞的子集识别为慢循环细胞。我们假设 存在特定的细胞表面标记物，可以定义和 选择角膜缘内的干细胞。此类标记的识别 将通过提供用于研究人类眼表的工具来帮助研究 两者都选择特定的药物或基因并将其靶向这些细胞。更远， 他们将提供用于分析角膜缘干细胞的工具 体内和体外的特性。此应用程序的具体目标 是： 1) 通过以下方法鉴定来自供体人角膜的角膜干细胞样细胞： 基于细胞粘附特性、整合素表达谱和 细胞体外增殖的能力，2.使用噬菌体展示肽 文库，鉴定与人类、小鼠特异性结合的肽， 和大鼠角膜缘基底细胞以及使用这些肽来分离和 表征干细胞特异性表面标记，3. 在小鼠和大鼠中， 确定新生儿后角膜缘是否观察到慢循环细胞 BrdU 给药具有与人类角膜相似的细胞表面标记 干细胞，以及 4. 使用目标 1-3 中开发的知识和工具， 表征小鼠的角膜缘干细胞在其 野生型和遗传型的分化和/或增殖发生改变 工程小鼠表现出干细胞疾病的症状。发现具体的 角膜缘干细胞的分子标记将有助于发展 治疗人类眼表干细胞缺陷性疾病。