Isolation of Limbal Epithelial Stem Cells

角膜缘上皮干细胞的分离

• 批准号: 6524804
• 负责人: ROBERT M LAVKER
• 金额: $ 14.65万
• 依托单位: NORTHWESTERN UNIVERSITY AT CHICAGO
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-08-01 至 2004-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6524804
• 关键词: Rodentias; biomarker; cell differentiation; cell growth regulation; cell population study; cell sorting; corneal epithelium; cytogenetics; gene expression; homeostasis; microarray technology; organ culture; single cell analysis; stem cells; subtraction hybridization

DESCRIPTION: (Applicant's Abstract) Epithelial stem cells are responsible for the homeostasis of self-renewing epithelia. Moreover they play a central role in wound repair and are the target cell for tumor initiation. My work during the past decade has contributed to the identification of stem cells within the corneal epithelium. Isolation of limbal epithelial stem cells will greatly facilitate their characterization. Furthermore, isolation of pure populations of limbal stem cells will provide the ultimate starting material for epithelial cells needed for transplantation in corneal reconstruction. The lack of specific markers for epithelial stem cells has become a major impediment in their isolation and subsequent biochemical characterization. The goal of this project is to identify specific markers for limbal epithelial stem cells. Towards this end we will: 1) use single cell technology in combination with molecular biology to obtain mRNA from limbal epithelial stem cells and corneal epithelial transit amplifying (TA) cells; 2) use suppressive subtractive hybridization (SSH) and cDNA microarrays to identify genes that are differentially expressed in limbal epithelial stem cells versus corneal epithelial TA cells; and 3) characterize the limbal epithelial stem cell-specific genes and investigate their potential function(s), focusing on those genes that are associated with the cell surface. Data obtained from these studies should lead to the isolation of pure populations of limbal epithelial stem cells. This will facilitate the biochemical characterization of these cells, and will provide a better understanding of the mechanisms involved in the self-renewal of stem cells, and in the mechanisms of limbal and corneal epithelial growth and differentiation.

描述：（申请人的摘要）上皮干细胞负责 自我更新上皮的稳态。而且他们扮演着核心角色 在伤口修复中，是肿瘤启动的靶细胞。我的工作期间 过去十年有助于鉴定干细胞 角膜上皮。层缘上皮干细胞的隔离将大大 促进他们的特征。此外，纯种群的隔离 边缘干细胞的最终起始材料的上皮 在角膜重建中需要移植所需的细胞。缺乏 上皮干细胞的特定标记已成为主要障碍 它们的隔离和随后的生化特征。目标的目标 项目是确定缘缘上皮干细胞的特定标记。 为此，我们将： 1）将单细胞技术与分子生物学结合使用 来自边缘上皮干细胞和角膜上皮转运的mRNA 扩增（TA）细胞； 2）使用抑制性减法杂交（SSH）和 cDNA微阵列鉴定在缘缘以差异表达的基因 上皮干细胞与角膜上皮TA细胞； 3）特征 边缘上皮干细胞特异性基因并研究其潜力 功能（s），重点关注与细胞表面相关的基因。 从这些研究获得的数据应导致纯 边缘上皮干细胞的种群。这将有助于 这些细胞的生化表征，将提供更好的 了解干细胞自我更新的机制，以及 在边缘和角膜上皮生长和分化的机制中。