SELF RENEWAL IN NORMAL & LEUKEMIC HEMOPOIETIC STEM CELLS

正常情况下的自我更新

• 批准号: 3167119
• 负责人: DONALD METCALF
• 金额: $ 8.22万
• 依托单位: WALTER AND ELIZA HALL INST MEDICAL RES
• 依托单位国家: 美国
• 财政年份: 1983
• 资助国家: 美国
• 起止时间: 1983-06-01 至 1989-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3167119
• 关键词: bioassay; bone marrow; cell differentiation; cell growth regulation; cellular oncology; chromatography; clone cells; genetic manipulation; granulocyte; growth media; hematopoietic stem cells; interleukin 3; leukocyte activation /transformation; leukopoietic factor; macrophage; molecular cloning; myelogenous leukemia; neoplasm /cancer transplantation; radiation leukemia; stromal cells; thymus; transposon /insertion element; viral leukemia

It is proposed to characterize active factors operating in recently developed methods for the in vitro culture and activation of pluripotential hemopoietic cells and to transfer DNA's for GM- CSF and MultiCSF into such cultured stem cells or their controlling stromal cells to determine the role of dysregulated CSF production in the development and progression of myeloid leukemia. The biochemical nature of the factors able to control the maintenance and expansion of pluripotential hemopoietic cell numbers will be determined using three types of starting materials (a) medium from the cloned B.Ad stromal cell line, (b) normal organ conditioned media (thymus, marrow shaft), and (c) U5637 conditioned medium. Biochemical fractionation will be monitored by three assay systems using post5FU bone marrow cells and will determine whether more than one such factor exists. Where appropriate, FACSpurified pluripotential cells will be used with purified recombinant CSF's to establish whether the stem cell-active factors have proliferative, activating or differentiation-inducing actions. DNA's for murine GM-CSF and Multi-CSF (IL-3) will be introduced (a) into hemopoietic stemm cells cultured with stimulation by the above factors, sing retroviral vectors and microinjection, then the cells used to repopulate irradiated recipients, and (b) into all tissues of the body by the generation of transgenic mice. The hemopoietic tissues from these contrasting mice will be analyzed to determine whether dysregulated CSF synthesis alone is sufficient for myeloid leukemia development or whether additional radiationinduced translations or the insertion of an additional oncgene are necessary.

提出了表征近期活动因素的方法 开发了体外培养和激活的方法， 多能造血细胞和转移DNA的GM- CSF和MultiCSF的干细胞或其组合。 控制基质细胞，以确定失调的 髓系白血病发生和发展过程中CSF的产生 白血病 生物化学性质的因素能够控制 多能造血细胞的维持和扩增 数字将使用三种类型的启动来确定 材料（a）来自克隆的B.Ad基质细胞系的培养基，（B） 正常器官条件培养基（胸腺、骨髓干），和（c） U 5637条件培养基。 生化分离将是 使用5 FU后骨髓，通过三种测定系统进行监测 单元格，并将确定是否有多个此类因素 存在. 在适当的情况下，FACS纯化的多能细胞将 与纯化的重组CSF一起使用，以确定 干细胞活性因子具有增殖、活化或 分化诱导行为。 鼠GM-CSF和Multi-CSF（IL-3）的DNA将 引入（a）培养的造血干细胞， 通过上述因子的刺激，单逆转录病毒载体和 显微注射，然后用于重新繁殖的细胞被照射 受体，和（B）通过产生 转基因小鼠 这些对比鲜明的造血组织 将分析小鼠以确定是否存在失调的CSF 单独的合成足以使骨髓性白血病发展，或者 无论是额外的辐射引起的平移还是插入 额外的致癌基因是必要的。