ANALYSIS OF LECTIN BINDING EPITOPES ON THE SURFACE OF MURINE ES CELLS

鼠 ES 细胞表面凝集素结合表位的分析

• 批准号: 7722617
• 负责人: Stephen Dalton
• 金额: $ 13.16万
• 依托单位: UNIVERSITY OF GEORGIA
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-08-08 至 2009-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7722617
• 关键词: Acetylgalactosamine; Alkaline Phosphatase; Binding; CD15 Antigens; Cell surface; Cells; Computer Retrieval of Information on Scientific Projects Database; Development; Ectoderm; Epitopes; Forssman Antigen; Funding; Grant; Institution; Kinetics; Lectin; Mus; Outcome; Polysaccharides; Population; Research; Research Personnel; Resolution; Resources; Source; Surface; United States National Institutes of Health; dolichos biflorus agglutinin; embryonic stem cell; tool

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Cell surface markers are key tools that are frequently used to characterize and separate mixed cell populations. Existing cell surface markers used to define murine embryonic stem cells (mESCs), such as stage specific embryonic antigen 1 (SSEA1), Forssman antigen (FA), alkaline phosphatase (AP) and CD9, are limiting, however, because they do not unambiguously define the pluripotent state and are not reliable indicators of differentiation commitment. To identify glycan cell surface markers that would circumvent this problem we used a panel of 18 lectins to identify epitopes specifically elevated on the surface of mESCs which, during differentiation, decrease with kinetics that precede currently used markers such as CD9, SSEA1, FA and AP. The anticipated outcome of this analysis was to identify glycans that have utility as a reliable mESC marker and as a high resolution readout for early differentiation commitment. Here we show that the lectin Dolichos biflorus agglutinin (DBA), recognizes alpha-N- acetylgalactosamine (GalNAc) cell surface epitopes on mESCs (CD9high SSEA1high APhigh DBAhigh). These glycan epitopes decline markedly in cells undergoing the first definable step of differentiation, the transition from mESCs to primitive ectoderm (CD9high SSEA1high APhigh DBAlow). Loss of GalNAc epitopes is therefore the earliest cell surface change that can be assigned to differentiating cells and the only cell surface marker known to be tightly associated with the pluripotent state. Therefore, the lectin DBA is a useful tool to characterize mESC cultures by non-destructive approaches, an indicator of differentiation commitment and a predictor of developmental potency.

该副本是利用众多研究子项目之一 由NIH/NCRR资助的中心赠款提供的资源。子弹和 调查员（PI）可能已经从其他NIH来源获得了主要资金， 因此可以在其他清晰的条目中代表。列出的机构是 对于中心，这不一定是调查员的机构。 细胞表面标记是经常用于表征和分离混合细胞种群的关键工具。 现有用于定义鼠类胚胎干细胞（MESC）的细胞表面标记，例如阶段特异性胚胎 然而，抗原1（SSEA1），Forssman抗原（FA），碱性磷酸酶（AP）和CD9是有限的，因为它们是限制的 不要明确定义多能状态，也不是分化承诺的可靠指标。到 识别可以避免此问题的聚糖细胞表面标记，我们使用了18个凝集素的面板来识别 在MESC表面上特别升高的表位，在分化过程中，随着动力学的降低， 目前使用了CD9，SSEA1，FA和AP等标记。该分析的预期结果是 识别具有效用作为可靠MESC标记的聚糖，并作为早期差异化的高分辨率读数 承诺。在这里，我们表明凝集素双纤维凝集素（DBA）认识到alpha-n-- MESC（CD9High ssea1 -High aphigh dbahigh）上的乙酰乳糖胺（GalNAC）细胞表面表面表现。这些聚糖 在经历第一个可确定分化步骤的细胞中，表位显着下降，从MESC到 原始外胚层（CD9High ssea1 -high aphigh dbalow）。因此，galnac表位的损失是最早的细胞 可以分配给分化细胞的表面变化，也是唯一已知紧密的细胞表面标记 与多能状态相关。因此，凝集素DBA是表征的有用工具 MESC培养通过非破坏性方法，分化承诺的指标和预测指标 发展效力。