CORE--STEM CELL AND TISSUE CULTURE

干细胞和组织培养核心将提供一个集中的设施 以及细胞系和造血干细胞纯化的能力， 计划项目中所有参与者的同步和文化。 具体地说，核心将提供集中的维护专业知识 细胞系的研究和对小鼠的纯化 造血干/祖细胞。这些净化将基于 细胞表面表型标记物的表达及其对Low的利用 罗丹明和Hoechst染色以确定原始静止细胞。 提纯程序将广泛使用目前的荧光 激活的细胞分选核心(BD FACS Star Plus)和一项新研究 目前正在开发的专用细胞分选核心将具有 来自Cytomation的MoFlo高速细胞分选机。后一个核心将 允许我们继续进行关于细胞的更多开发工作 分类，并将允许更快的分类和实验 白天和晚上的几个小时。CORE还将开发和完善协议 用于祖细胞干细胞同步和移植。通过 将细胞培养活动整合到单一来源中，核心将 标准化细胞群的功能，由各种不同的 并将能够为各个项目提供统一的 细胞因子和培养试剂的质量和持续供应。一个 核心的关键特征将是提供高度净化的 每个项目的祖细胞/干细胞群体和 开发新的移植模式。

The Stem Cell and Tissue Culture Core will provide a centralized facility and capacity for cell line and hematopoietic stem cell purification, synchronization and culture for all participants in the Program Project. Specifically, the Core will provide a focused expertise in the maintenance of cells lines and a major emphasis on the purification of murine hematopoietic stem/progenitor cells. These purifications will be based on cell surface expression of phenotypic markers and the utilization of low staining with Rhodamine and Hoechst to define primitive quiescent cells. Purification procedures will make extensive use of the current fluorescent activated cell sorting core (BD FACS Star plus) and of a new research dedicated cell sorting core currently under development which will feature the MoFlo high speed cell sorter from Cytomation. This latter core will allow us to proceed with more developmental work with regard to cell sorting and will allow for higher speed sorting and experiments at all hours of the day and night. Cores will also develop and refine protocols for progenitor stem cell synchronization and transplantation. By consolidating cell culture activities in a single source the core will function ot standardized cell populations used in parallel by the various projects and will be able to provide individual projects with a uniform quality and consistent supply of cytokines and culture reagents. A critical feature of the core will be the provision of highly purified progenitor/stem cell populations to each of the projects and the development of new transplantation models.