Purification and ampliefication of hematopoietic stem cells

造血干细胞的纯化和扩增

• 批准号: 06277104
• 负责人: SUDA Toshio
• 金额: $ 128.83万
• 依托单位: Kumamoto University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research on Priority Areas
• 财政年份: 1994
• 资助国家: 日本
• 起止时间: 1994 至 1998
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-06277104/
• 关键词: hemotopoietic stem cells; receptor tyroshine kinases; CD34; IL-6R; GATA; 造血前駆細胞; シグナル伝達; 血液幹細胞; AGM領域; 細胞運命決定因子; 自己複製能; 多分化能; 転写調節; 赤血球分化; gp130; 蛍光細胞分離装置; IL-6; 幹細胞増幅; 転写因子

Soda, Miura and Kanakura have shown that some signal transduction pathways in hemopoictic stem cells (HSC). Novel receptor tyrosine kinases such as TIEs, HTK and STK/RON were cloned from purified HSC by RT-PCR and especially TIE-1 and 2 are expressed in hematopotetic stem cells as well as in vascular endothelial cells From these findings, it becomes possible to analyze the development of hematopoietic stem cells from angioblasts, which are endothelial precursor cells. It was clarified that gain of function of c Kit caused the autonomous proliferation of hematopoietic cells (mast cells) and specified cells in the gut (Kajal cells).Nakauchi has demonstrated that CD34tィイD1+ィエD1KitィイD1+ィエD1ScaィイD1+ィエD1LinィイD1-ィエD1 and CD34ィイD1+ィエD1KitィイD1+ィエD1ScaィイD1+ィエD1LinィイD1-ィエD1 cells of adult murine bone marrow have been shown to contain long-and short-term repopulating hematopoictic stem cells by a transplantation assay. CD34 negative stein cells are also confirmed in human hematopoletic system. There has been great interest in the ex vivo expansion of human hemopoictic stem/progenitor cells for clinical applications. Nakahata has shown that heanatopoietic progenitor cellscan be expanded in the presence of soluble IL-6 receptor+IL-6+SCF. However. stein cell expansion is still insatisfactoiy. Asano has tried to establish monkey transplantation system to analyze the hurtan HSC.Yamamoto has clearly shown that GATA and Maf families am involved in erythropoiesis by the GATA-1 KO mice and GATA promoter-LacZ transgenic mice.

Soda、Miura和Kanakura已经证明了造血干细胞(HSC)中的一些信号转导途径。通过RT-PCR从纯化的HSC中克隆到了TIES、HTK和STK/RON等新的受体酪氨酸激酶，特别是Tie-1和Tie-2在造血干细胞和血管内皮细胞中都有表达，这些发现为分析血管母细胞作为内皮前体细胞的造血干细胞的发育提供了可能。研究表明，c-Kit功能的增强导致了造血细胞(肥大细胞)和肠道特定细胞(Kajal细胞)的自主增殖。中内已通过移植实验证明，成年小鼠骨髓的CD34tィイD1+ィエD1KitィイD1+ィエD1ScaィイD1+ィエD1LinィイD1-ィエD1和CD34ィイD1+ィエD1KitィイD1+ィエD1ScaィイD1+ィエD1LinィイD1-ィエD1细胞含有长期和短期的再生造血干细胞。在人类造血系统中也证实了CD34阴性的Stein细胞。人类造血干/祖细胞的体外扩增在临床上的应用引起了人们的极大兴趣。Nakahata已经证明，在可溶性IL-6受体+IL-6+SCF存在的情况下，造血祖细胞可以被扩增。然而。Stein细胞的扩增仍然不令人满意。Asano等曾尝试建立猴移植系统来分析人HSC，Yamamoto已经明确表明GATA和Maf家族参与了GATA-1 KO小鼠和GATA启动子-LacZ转基因小鼠的红细胞生成。

项目成果

HiraO A: "Translocation of the Csk homologous kinase(Chk/Hyl)controls activity of CD36-anchored Lyn tyrosine in thrombin-stimulated platelets." EMBO J. 16. 2342-2351 (1997)

HiraO A：“Csk 同源激酶 (Chk/Hyl) 的易位控制了凝血酶刺激的血小板中 CD36 锚定的 Lyn 酪氨酸的活性。”

Iwama A et al.: "Molecular cloning of a novel receptor tyrosine kinase gene,STK,derived from enriched hematopoietic stem cells." Blood. 83. 3160-3169 (1994)

Iwama A 等人：“源自富集造血干细胞的新型受体酪氨酸激酶基因 STK 的分子克隆。”

Itoh K et al: "Cloning and characterization of a novel erythroid-derived CNC family transcription factor heterodimerizing with the small Maf." Mol.Cell.Biol. 15. 4184-4193 (1995)

Itoh K 等人：“与小 Maf 异二聚化的新型红系衍生 CNC 家族转录因子的克隆和表征。”

Komatsu N et al: "In vitro development of erythrold and megakaryocytic cells from a UT-7 subline, UT-7/GM"Blood. 89. 4021-4033 (1997)

Komatsu N 等人：“来自 UT-7 亚系 UT-7/GM 的红细胞和巨核细胞的体外发育”血液。

Matsuzaki Y.: "Role of bcl-2 in the Development of Lymphoid Cells From the Hematopoietic Stem Cell." Blood. 89. 853-862 (1997)

Matsuzaki Y.：“bcl-2 在造血干细胞发育成淋巴细胞中的作用。”