CORE--CELL SEPARATION

核心--细胞分离

基本信息

批准号：
6494852
负责人：
ROBERT GOFF ANDREWS
金额：
$ 20.94万
依托单位：
FRED HUTCHINSON CANCER RESEARCH CENTER
依托单位国家：
美国
项目类别：
财政年份：
2001
资助国家：
美国
起止时间：
2001-09-01 至 2002-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/6494852
关键词：
animal tissue biomedical facility flow cytometry hematopoietic stem cells human tissue tissue /cell culture

项目摘要

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.

使用流式细胞术纯化正常造血祖细胞和干细胞，以及分析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，这是本次资助项目的核心。流式细胞术可用于根据细胞表面和细胞内决定簇的表达、物理特性（通过光散射确定细胞大小）以及活体染料标记来分析和纯化活细胞和固定细胞。流式细胞术还可用于研究细胞表面决定簇和细胞内分子（包括蛋白质和 mRNA）的同时表达，以及用于细胞周期分析的 DNA 含量。细胞分离核心将使用流式细胞术执行所有细胞分析和细胞分选，包括批量和单细胞沉积（克隆）。本次资助的每个项目都将使用流式细胞术来纯化祖细胞和干细胞，用于直接体外和体内研究，并研究干细胞及其后代的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以及培养和感染的原代细胞进行多参数流式细胞术分析，以及多参数细胞分选和单细胞克隆，以分离人类、小鼠和狒狒骨髓和血液的子集细胞以及体外所需的干细胞和祖细胞。