CORE--CONTROL OF STEM CELL DIFFERENTIATION IN THE THYMUS

描述（直接取自应用程序）：了解 机制 造血干细胞功能需要体外谱系分析 承诺.对于T谱系，这仅可通过离体胸腺移植获得。 器官培养我们的目标是提供我们记录在案的专业知识， 技术研发人员。此外，我们计划优化 并扩展这项技术，使其对未来的T 血统承诺 I.我们将进行胸腺器官培养试验，以评估T淋巴细胞 造血干细胞的潜能。我们的实验室优化了 标准的胸腺器官培养技术，目前正在开发 提高胸腺器官实验利用率的新途径 文化例如，我们开发了一种逆转录病毒技术， 标记将允许祖细胞的克隆分析。我们将 进行胎儿胸腺器官重建，此外，限制 稀释测定和胸腺再聚集研究。 二.我们将提供试剂和专业知识，用于评估T 体内模型系统中的淋巴发育和功能。T谱系 发展的特点是各种各样的表达变化 形态学标记。我们将使用抗体染色和免疫组织化学方法， 流式细胞术分析以评估T细胞的分化状态及其对免疫应答的影响。 祖先此外，T细胞功能将通过体内试验进行评估。 以及体外刺激，随后进行细胞因子产生、T细胞 增殖和细胞毒性。 三.当前和未来的项目旨在评估特定的 造血谱系定型的基因。我们将提供载体，细胞 株系、细胞系和逆转录病毒介导基因的技术支持 造血干细胞和淋巴祖细胞的转移，以及 基质，用于胸腺中外源基因的表达。

Description (taken directly from the application): Understanding the mechanism of hematopoietic stem cell function requires in vitro analysis of lineage commitment. For the T lineage, this is only available by ex vivo thymic organ culture. Our goal is to provide our documented expertise in this technology to Program Investigators. In addition, we plan to optimize and extend this technology to make it more useful for future studies of T lineage commitment. I. We will perform thymic organ culture assays to evaluate the T lymphoid potentials of hematopoietic stem cells. Our laboratory has optimized standard thymic organ culturing techniques, and is currently developing new approaches to increase the experimental utility of thymic organ culture. For example, we have developed a technique for retroviral marking that will allow the clonal analysis of progenitors. We will perform fetal thymic organ reconstitutions, and in addition, limiting dilution assays and thymic reaggregation studies. II. We will provide reagents and expertise for the evaluation of T lymphoid development and function in vivo model systems. T lineage development is characterized by changes in expression of a wide variety of morphological markers. We will use antibody staining and multicolor FACS analysis to assess the differentiative status of T cells and their progenitors. In addition, T cell function will be evaluated by in vivo and in vitro stimulation followed by tests of cytokine production, T cell proliferation, and cytotoxicity. III. Current and future projects aim to evaluate the function of specific genes in hematopoietic lineage commitment. We will provide vectors, cell lines, cell lines, and technical support for retrovirus-mediated gene transfer in hematopoietic stem cells and lymphoid progenitors, and stroma, for expression of exogenous genes in thymus.