CLONAL HEMATOPOIETIC PROGENITOR CELL ASSAY

克隆造血祖细胞测定

• 批准号: 3502482
• 负责人: MANFRED R KOLLER
• 金额: $ 5万
• 依托单位: AASTROM BIOSCIENCES, INC.
• 依托单位国家: 美国
• 财政年份: 1993
• 资助国家: 美国
• 起止时间: 1993-08-01 至 1994-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3502482
• 关键词: bone marrow; cell sorting; clone cells; growth media; hematopoiesis; hematopoietic stem cells; human subject; method development; microcapsule; perfusion; single cell analysis; tissue /cell culture

Hematopoiesis, the regulated production of mature blood cells, is a complex scheme of multilineage differentiation which occurs mainly in the bone marrow of adult mammals. Mature cells are continuously produced from progenitor (or percursor) cells. Consequently, assays which detect these progenitor cells are vital in many research and clinical settings. The current assay procedures suffer from a number of shortcomings. First, they are carried out under suboptimal conditions which do not represent the in vivo environment. As a result, the assays are slow and the results may not be physiologically relevant. Furthermore, many current assay techniques do not ensure clonality, thereby precluding the harvest and study of the progeny of a single cell. Finally, most assays allow accessory cell interactions, and this often obscures the true action of the activity being assayed. These drawbacks can be overcome by the use of microencapsulation and rapid perfusion. We propose a Phase I study whose primary objective is to demonstrate that single progenitor cells can be grown out clonally in a microcapsule. If this ability is demonstrated, then a Phase II study would follow during which the detailed dynamics of outgrowth, including kinetics, lineage reconstitution, and expansion of immature hematopoietic cells, would be established. This assay would then be made available on a contract basis for growth factor screening, drug efficacy testing, toxicology studies and as a radiation assay.

造血，成熟血细胞的调节生产，是一种 多谱系分化的复杂方案，主要发生在 成年哺乳动物的骨髓成熟的细胞不断地从 祖细胞（或前体细胞）。因此，检测这些的测定法， 祖细胞在许多研究和临床环境中至关重要。的 目前的分析方法存在许多缺点。一是 是在次优条件下进行的， 体内环境因此，测定缓慢，结果可能不 与生理相关。此外，许多当前的测定技术不 不能确保克隆性，从而排除了收获和研究 单细胞的后代。最后，大多数测定允许辅助细胞 交互，这往往掩盖了活动的真实作用 正在化验。这些缺点可以通过使用 微囊化和快速灌注。我们提出了一项I期研究， 主要目的是证明单个祖细胞可以 在一个微胶囊里克隆生长如果这种能力被证明， 然后进行第二阶段研究，在此期间， 副产物，包括动力学，谱系重建和扩张， 不成熟的造血细胞，将被建立。然后，该测定将 在合同的基础上提供生长因子筛查，药物 功效测试、毒理学研究和作为辐射分析。