DNA ARRAY ANALYSIS OF STEM CELL DIFFERENTIATION II

描述(申请人提供)：糖蛋白和糖脂在人类生理和疾病中具有多种功能，这些细胞表面分子上的糖链的表达在早期胚胎发育过程中会发生变化。来自早期胚胎的胚胎干细胞可以在体外培养并诱导分化为多种细胞类型。控制这种分化并产生同质的分化干细胞群体是生物医学界面临的主要挑战。细胞标记物的发展和丰富细胞群体的技术可以极大地帮助干细胞研究。我们建议建立一个生物医学糖学综合资源中心，专注于开发工具和技术，详细分析小鼠胚胎干细胞及其分化为细胞的糖蛋白和糖脂表达。项目1，胚胎干细胞计划，将开发培养大量干细胞的工具，并详细研究特定细胞表面糖蛋白上表达的多糖。项目2将开发技术来分离干细胞产生的糖肽，并确定表达这些糖链的亲本蛋白以及这些糖链表达的位点。项目3将发展动态实时聚合酶链式反应来研究糖基转移酶转录本的表达。项目4将开发生物信息学工具，将结构和表达数据输入关系数据库进行处理和分析。在该资源中开发的技术将使人们能够了解糖基化在分化过程中是如何控制的，并将允许开发工具来促进使用干细胞治疗人类疾病。此外，这项技术将适用于其他细胞类型的研究，包括正在向更具侵袭性表型发展的癌细胞。这种糖组分资源将导致技术开发，使科学界的其他人能够通过传播开发的新方法和通过该资源向其他需要帮助进行糖组分分析的科学家提供分析服务来参与糖组分研究。还将向科学界提供有关已开发技术的培训课程。合作项目已经构成了拟议研究的大量内容，并将成为正在开发的糖组分资源中日益重要的组成部分。

DESCRIPTION (provided by applicant): Glycoproteins and glycolipids have many functions in human physiology and disease and the expression of the glycans on these cell surface molecules are known to change during early embryonic development. Embryonic stem cells, derived from early embryos, can be grown in vitro and induced to differentiate into many cell types. Controlling this differentiation and producing homogeneous populations of differentiated stem cells are major challenges for the biomedical community. Development of cell markers and technology to enrich cell populations could aid significantly stem cell research. We propose to establish an Integrated Resource Center for Biomedical Glycomics that will focus on developing tools and technology to analyze in detail the glycoprotein and glycolipid expression of mouse embryonic stem cells and cells into which they differentiate. Project 1, Embryonic Stem Cell Program, will develop the tools to culture large numbers of stem cells and investigate in detail the glycans expressed on specific cell surface glycoproteins. Project 2 will develop technology to fractionate glycopeptides produced from stem cells and determine the parent proteins that express these glycans and at which sites these glycans are expressed. Project 3 will develop kinetic real-time PCR to study the expression of glycosyltransferase transcripts. Project 4 will develop Bioinformatic tools to input structural and expression data into relational databases for processing and analysis. The technology developed in this Resource will allow an understanding of how glycosylation is controlled during differentiation and will allow the development of tools to promote the use of stem cells to treat human disease. In addition, the technology will be applicable to the study of other cell types, including cancer cells that are progressing to a more invasive phenotype. This glycomics resource will result in technology development that will allow others in the scientific community to participate in glycomics research via dissemination of the new methods developed and via analytical service provided by the resource to other scientists requesting help in glycomic analyses. Training courses in the developed technologies will also be provided to the scientific community. Collaborative projects already constitute a significant amount of the proposed research and will become an increasingly important component of the developing glycomics resource.