Characterization of Corneal Epithelial Stem Cells

角膜上皮干细胞的表征

• 批准号: 6360365
• 负责人: Mary Ann Stepp
• 金额: $ 32.38万
• 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/6360365
• 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)：本项目的长期目标是 使用分子方法定义一组细胞特异性表面受体 人类和啮齿动物的角膜缘干细胞，并将这些标记用作工具 人、大鼠和小鼠角膜上皮干细胞的特性 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细胞的来源。 目前，实验证实，角膜缘中的细胞是干细胞 依赖于体外细胞增殖能力研究的数据和 将这些细胞的一个子集识别为慢周期细胞。我们假设 存在一个特定的细胞表面标记谱系，它可以定义和 选择角膜缘内的干细胞。此类标记的识别 将通过提供工具来帮助研究人类眼表 两者都选择并靶向这些细胞的特定药物或基因。此外， 他们将提供分析角膜缘干细胞的工具。 体内和体外的性质。本申请的具体目的 方法：1)从供体人角膜中鉴定角膜干细胞 基于细胞黏附特性、整合素表达谱和 细胞的体外增殖能力，2.用噬菌体展示的多肽 文库，鉴定与人、鼠、 和大鼠角膜缘基底细胞，并使用这些肽来分离和 鉴定干细胞特异性表面标记，3.在小鼠和大鼠中， 确定新生鼠角膜缘是否存在慢周期细胞 BrdU给药具有与人类角膜相似的细胞表面标记 4.利用AIMS 1-3中开发的知识和工具， 在小鼠角膜缘干细胞的条件下鉴定其特性 分化和/或增殖在野生型和遗传型中发生改变 表现出干细胞疾病症状的转基因小鼠。发现特定的 角膜缘干细胞的分子标记将有助于角膜缘干细胞的发展 人类眼表干细胞缺乏症的治疗。