Characterization of Corneal Epithelial Stem Cells

角膜上皮干细胞的表征

• 批准号: 6794662
• 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/6794662
• 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 使用分子方法来定义一组细胞特异性表面标记物， 人类以及啮齿动物角膜缘干细胞，并使用这些标记物作为工具， 在人类、大鼠和小鼠中表征角膜上皮干细胞， 健康和疾病。临床经验和实践表明，角膜缘 干细胞作为构成角膜上皮的细胞的来源。 目前，实验证实，在利姆布斯细胞是干细胞 依赖于来自体外细胞增殖能力研究的数据， 将这些细胞的子集识别为慢循环。我们假设 存在细胞表面标志物的特定库，其可以定义和 选择角膜利姆布斯内的干细胞。识别这些标记 将通过提供工具来帮助研究人眼表面， 两者都选择并将特定的药物或基因靶向这些细胞。此外，本发明还 他们将为角膜缘干细胞分析提供工具 在体内和体外的特性。本申请的具体目的是 是：1）从供体人角膜中鉴定角膜干细胞样细胞， 基于细胞粘附特性、整联蛋白表达谱和 细胞在体外增殖的能力，2.利用噬菌体展示肽 文库，特异性结合人，小鼠， 和大鼠角膜缘基底细胞以及这些肽用于分离和 表征干细胞特异性表面标志物，3.在小鼠和大鼠中， 确定新生儿后在利姆布斯中观察到的慢循环细胞是否 BrdU给药具有与人角膜相似的细胞表面标志物 干细胞，以及4.利用在目标1-3中开发的知识和工具， 表征小鼠角膜缘干细胞的条件下， 分化和/或增殖在野生型中和在遗传学上是改变的。 表现出干细胞疾病症状的工程小鼠。发现具体 角膜缘干细胞的分子标记将有助于 治疗人类眼表干细胞缺乏性疾病。