CORE--CELL SEPARATION

使用流式细胞术纯化正常造血祖细胞和干细胞，以及分析由造血细胞表达的特定细胞表面和细胞内分子的表达和功能，作为本次赠款项目的中心。流式细胞术可用于分析和纯化活的和固定的细胞，基于细胞表面和细胞内决定因素的表达，物理性质(细胞大小由光散射确定)，以及活性染料的标记。流式细胞术还可以用来研究细胞表面决定因素和细胞内分子(包括蛋白质和mRNA)的同时表达，以及细胞周期分析中的DNA含量。细胞分离核心将使用流式细胞术进行所有细胞分析和细胞分类，包括批量和单个细胞沉积(克隆)。这笔赠款中的每个项目都将使用流式细胞术来纯化祖细胞和干细胞，用于直接进行体外和体内研究，并研究干细胞及其后代细胞表面和细胞内分子的表达。因此，细胞分离核心设施的功能将是对细胞系和培养和感染的原代细胞进行多参数流式细胞仪分析，以及多参数细胞分选和单细胞克隆，以分离体外所需的人、小鼠和狒狒骨髓和血细胞亚群以及干细胞和祖细胞。

The use of flow cytometry for the purification of normal hematopoietic progenitor and stem cells as well as to analyze the expression and function of defined cell surface and intracellular molecules expressed by hematopoietic cells as central to the projects in this grant. Flow cytometry can be used to analyze and purify viable and fixed cells based on the expression of cell surface and intracellular determinants, physical properties (cell size determined by light scattering), and labeling with vital dyes. Flow cytometry also can be used to study the simultaneous expression of cell surface determinants and intracellular molecules including proteins and mRNA, as well as DNA content for cell cycle analysis. The Cell Separation Core will perform all cell analysis and cell sorting, for both bulk and single cell deposition (cloning), using flow cytometry. Flow cytometry will be used by each of the projects in this grant to purify progenitor and stem cells for direct in vitro and in vivo studies and to study expression of cell surface and intracellular molecules by stem cells and their progeny. Therefore, the function of the Cell Separation Core facility will be to perform multiple parameter flow cytometric analysis of cell lines and cultured and infected primary cells and multi-parameter cell sorting and single cell cloning to isolate subsets of human, mouse and baboon marrow and blood cells as well as stem and progenitor cells necessary for in vitro.