Studies of differentiation mechanisms of hematopoietic stem cells using cytokine-receptor transgenic mice

利用细胞因子受体转基因小鼠研究造血干细胞分化机制

基本信息

批准号：
10307020
负责人：
NAKAHATA Tatsutoshi
金额：
$ 24.83万
依托单位：
KYOTO UNIVERSITY (1999)The University of Tokyo (1998)
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (A)
财政年份：
1998
资助国家：
日本
起止时间：
1998 至 1999
项目状态：
已结题

项目摘要

Granulocyte-macrophage colony stimulating factor (GM-CSF) stimulates proliferation and maturation of myeloid progenitor cells through cell surface receptors expressed on target cells. To analyze the relationship between receptor expression and differentiation of hemopoietic cells, transgenic mice (Tg mice) which express hGMR at all stages of hemopoitic cell development were generated using hGMR α and β cDNAs. hGM-CSF supported colony formation of various types including blast cell, erythroid (E), megakaryocyte (Meg), and mixed hematopoietic colonies. Administration of hGM-CSF to Tg mice resulted in increase of not only neutrophils, monocyte, eosinophils, NK cells but also erythrocytes in their peripheral blood. To clalify whether GM-CSFR signaling can compensate erythropoietin receptor (EPOR) signaling in erythropoiesis, we next generated double mutant mice, hGM-CSFR+/+ and EPOR-/-. Although double mutant mice were embryonic lethal, hGM-CSF induced a large number of erythroid colonies from day 14 fetal liver cells of double mutant mice. We also generated Tg mice that had ubiquitous expressions of wild type human G-CSFR, wild type murine G-CSFR. In methylcellulose colony assay of bone marrow and spleen cells G-CSF have an effect on promoting the proliferation and differentiation of not only granulocyte but also macrophage, megakaryocyte, mast cells, erythroid and more primitive hematopoietic progenitors in both serum-containing and -free culture. These results shows that when functional GM-CSFR or G-CSFR are present on the cell surface, both cytokines does not induce exclusive commitment to the neutrophil and macrophage lineages. In vitro and in vivo studies using the Tg mice indicated that cytokines such as hGM-CSF or hG-CSF supported the growth of various hematopoietic progenitors when the receptor was expressed, but did not alter their commitment program, thus favoring the "stochastic model" rather than the "deterministic model".

粒细胞巨噬细胞集落刺激因子（GM-CSF）刺激髓样祖细胞通过在靶细胞上表达的细胞表面受体的增殖和成熟。为了分析受体表达与血态细胞分化之间的关系，使用HGMRα和βcDNA产生了在血有质细胞发育的所有阶段表达HGMR的转基因小鼠（TG小鼠）。 HGM-CSF支持各种类型的菌落形成，包括爆炸细胞，红细胞（E），巨核细胞（MEG）和混合造血菌落。 HGM-CSF对TG小鼠的施用不仅会增加中性粒细胞，单核细胞，嗜酸性粒细胞，NK细胞，而且还增加了外周血中的红细胞。 To clalify whether GM-CSFR signaling can compensate erythropoietin receptor (EPOR) signaling in erythropoiesis, we next generated double mutant mice, hGM-CSFR+/+ and EPOR-/-.Although double mutant mice were embryonic lethal, hGM-CSF induced a large number of erythroid colonies from day 14 fetal liver cells of double mutant mice.We also generated Tg mice that had ubiquitous expressions of wild type human G-CSFR, wild type murine G-CSFR.In methylcellulose colony assessment of bone marrow and spleen cells G-CSF have an effect on promoting the proliferation and differentiation of not only granulocyte but also macrophage, megakaryocyte, mast cells, erythroid and more primitive hematopoietic progenitors in both含血清和无血清培养。这些结果表明，当功能性GM-CSFR或G-CSFR存在于细胞表面时，两种细胞因子都不会引起对中性粒细胞和巨噬细胞谱系的独家承诺。使用TG小鼠的体外和体内研究表明，当表达受体时，HGM-CSF或HG-CSF等细胞因子（例如HGM-CSF或HG-CSF）支持各种造血祖细胞的生长，但并没有改变其承诺程序，因此有利于“随机模型”而不是“确定性模型”。