SELF RENEWAL IN NORMAL & LEUKEMIC HEMOPOIETIC STEM CELLS

正常情况下的自我更新

• 批准号: 3167123
• 负责人: DONALD METCALF
• 金额: $ 7.94万
• 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/3167123
• 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; laboratory mouse; 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.

建议用来刻画近期运行的活跃因素 建立了体外培养和激活的方法 多能造血细胞和转移DNA用于GM- 将脑脊液和多核细胞集落刺激因子导入此类培养的干细胞或其 控制基质细胞以确定失调的作用 髓系细胞发生发展过程中脑脊液的产生 白血病。 能够控制的因素的生化性质 多潜能造血细胞的维持和扩增 数字将使用三种类型的起始值确定 材料(A)来自克隆的B.Ad基质细胞系的培养液，(B) 正常器官条件培养液(胸腺、骨髓干)，和(C) U5637条件培养液。生化分馏将是 使用Post 5FU骨髓的三种检测系统监测 单元格，并将确定是否有一个以上这样的因素 是存在的。在适当的情况下，FAC纯化的多潜能细胞将 与纯化的重组脑脊液一起用于确定 干细胞活性因子具有增殖、活化或 分化诱导作用。 小鼠GM-CSF和多粒细胞集落刺激因子(IL-3)的DNA将是 将(A)导入体外培养的造血干细胞 上述因素的刺激，逆转录病毒载体和 显微注射，然后用照射后的细胞重新填充 受体，以及(B)通过产生 转基因小鼠。来自这些对比鲜明的组织的造血组织 将对小鼠进行分析，以确定脑脊液调节失调 仅合成就足以发展成髓系白血病或 无论是额外的辐射诱导的翻译或插入 一个额外的致癌基因是必要的。