CORE-- STEM CELL SUPPORT

核心——干细胞支持

• 批准号: 7077699
• 负责人: Peter I Oettgen
• 金额: $ 7.94万
• 依托单位: BETH ISRAEL DEACONESS MEDICAL CENTER
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/7077699
• 关键词: biological models; biomedical facility; cell differentiation; flow cytometry; laboratory mouse; microarray technology; myocardial infarction; myocardial ischemia /hypoxia; stem cell transplantation; stem cells; vascular endothelium

DESCRIPTION (provided by applicant): The core will function as a service facility to investigators interested in using mouse models of ES cell differentiation to evaluate endothelial-specific gene expression during the process of endothelial differentiation. The in vitro differentiation of embryonic stem cells is a classic model of cellular differentiation that recapitulates many of the events that occur during normal embryonic development. A mayor focus of this Program Project is the identification of molecular mechanisms underlying endothelial specific gene expression. The differentiation of murine ES cells into embyroid bodies will serve as a model system for defining the molecular mechanism required for endothelial-specific gene expression in the developing embryo. This model has been well characterized and recapitulates many of the events that occur during normal mouse embryogenesis. Reproducible sequential expression of endothelial specific genes accompanied by the formation of endothelial cells and primitive vascular structures can be observed in this model by day 10 of differentiation. This model will allow the determination of which regulatory elements from the endothelial-specific genes, using Hprt-targeted constructs employing selected regulatory elements from the endothelial-specific genes to direct LacZ expression. In addition, the embryoid model will serve to identify novel surface markers that mark the transition from an undifferentiated ES cell toward a fully differentiated endothelial cell and facilitate the identification of better markers of endothelial progenitor cells. Specific subsets of cells will be separated by flow cytometry, allowing the identification of additional genes enriched in subsets of cells by microarray analysis, including the identification of potential novel transcriptional regulators of endothelial differentiation. Finally, the isolation and separation of particular subsets of cells of the endothelial lineage can be tested for their ability to promote angiogenesis in animal models of myocardial ischemia and infarction.

描述（由申请人提供）： 该核心将作为一个服务设施，有兴趣使用小鼠ES细胞分化模型，以评估内皮细胞分化过程中的内皮特异性基因表达的研究人员。胚胎干细胞的体外分化是细胞分化的经典模型，其概括了在正常胚胎发育期间发生的许多事件。该项目的主要重点是确定内皮特异性基因表达的分子机制。小鼠ES细胞分化为胚状体将作为一个模型系统，用于定义在发育胚胎中内皮特异性基因表达所需的分子机制。该模型已得到很好的表征，并概括了正常小鼠胚胎发生过程中发生的许多事件。在分化第10天，在该模型中可以观察到内皮特异性基因的可再现的连续表达，伴随着内皮细胞和原始血管结构的形成。该模型将允许使用Hprt靶向构建体来确定哪些调控元件来自内皮特异性基因，所述Hprt靶向构建体采用来自内皮特异性基因的选定调控元件来指导LacZ表达。此外，胚状体模型将用于鉴定新的表面标志物，其标记从未分化的ES细胞向完全分化的内皮细胞的转变，并促进内皮细胞的分化。 鉴定更好的内皮祖细胞标志物。特定的细胞亚群将被 通过流式细胞术分离，允许通过微阵列分析鉴定在细胞亚群中富集的另外的基因，包括鉴定内皮分化的潜在的新型转录调节因子。最后，可以在心肌缺血和梗塞的动物模型中测试内皮谱系细胞的特定亚群的分离和离析促进血管生成的能力。