Bone Marrow Microenvironments for Hematopoiesis

造血的骨髓微环境

• 批准号: 15590344
• 负责人: HAYASHI Shin-ichi
• 金额: $ 2.3万
• 依托单位: National University Corporation Tottori University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2003
• 资助国家: 日本
• 起止时间: 2003 至 2004
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-15590344/
• 关键词: Bone marrow; Hematopoiesis; Osteoclast; B cell; Toll-like receptor; Lipopolysaccharide(LPS); Tyrosine phosphatase; Embryonic stem(ES) cell; 髄外造血; 幹細胞; 造血支持細胞; Notchシグナル; リポポリサッカライド

1)Analysis of osteoclastogenesis for bone marrow formation :Osteoclasts are hematopoietic cells, which participate in bone resorption and remodeling. Receptor activator of NF-κB ligand(RANKL) and macrophage colony-stimulating factor(M-CSF) are critical for development of osteoclasts. The Toll-like receptor(TLR) family shares some of the downstream signaling with RANK, however, the signaling via TLRs has never been reported to induce osteoclastogenesis without RANKL. We showed that significant numbers of mature osteoclasts were generated from protein tyrosine phosphatase SHP-1-defective me^v/me^v bone marrow cells in the presence of M-CSF and LPS, a TLR4 ligand without addition of RANKL in culture. The osteoclast precursors were present in the Kit-positive cell enriched fraction of BM cells. Although me^v/me^v bone marrow cells required a comparable concentration of RANKL or TNF-α as wild-type cells for the initiation of osteoclastogenesis, numbers of multinucleated osteoclasts in me^v/ … More me^v bone marrow cultures were significantly increased by the equivalent dose of RANKL or TNF-α in the presence of M-CSF. These results indicate that a defect of SHP-1 function not only accelerates physiological osteoclast development by RANKL/RANK, but also acquires an aberrant pathway for osteoclastogenesis by LPS.2)Induction of hematopoiesis from blood-less embryonic stem (ES) cell line :Transcription factor Tal-1 is essential for the specification of hematopoietic development. Mice lacking Tall fail to generate any hematopoietic precursors. Using our co-culture system with stromal cells, we demonstrate that enforced expression of the transcription factor PU.1 under tetracycline control in Tall-null ES cells rescues the development of osteoclasts and macrophage-like phagocytes expressing a macrophage restricted marker. Other hematopoietic lineage cells were not generated. Their development was dependent on M-CSF and RANKL. These results suggest that the expression of PU.1 is a critical event for osteoclastogenesis and that Tal-1 may lie upstream of PU.1 in a regulatory hierarchy during osteoclastogenesis. Less

1)破骨细胞在骨髓形成中的作用分析：破骨细胞是造血细胞，参与骨吸收和改建。核因子受体激活剂B配体(RANKL)和巨噬细胞集落刺激因子(M-κ)对破骨细胞的发育起着至关重要的作用。Toll样受体(Toll-like Receptor，TLR)家族与RANK家族有一些共同的下游信号，然而，在没有RANKL的情况下，通过TLRs诱导破骨细胞生成的信号尚未见报道。我们发现蛋白酪氨酸磷酸酶SHP-1缺陷的Me、v/Me、v骨髓细胞在M-CSF和不加RANKL的TLR4配体的作用下，可以产生大量成熟的破骨细胞。破骨细胞前体存在于BM细胞的Kit阳性细胞富集区。尽管Me^v/Me^v骨髓细胞需要与野生型细胞相同浓度的RANKL或肿瘤坏死因子-α来启动破骨细胞生成，但在Me^v/…中多核破骨细胞的数量等剂量的RANKL或肿瘤坏死因子-α在M-α的存在下可显著增加骨髓培养的数量。这些结果表明，SHP-1功能缺陷不仅通过RANKL/RANK促进生理性破骨细胞的发育，而且通过LPS获得破骨细胞生成的异常途径。2)无血胚胎干细胞系的造血诱导：转录因子Tal-1对造血发育的规范是必不可少的。缺乏TALL的小鼠不能产生任何造血祖细胞。使用我们的基质细胞共培养系统，我们证明了在四环素控制下，转录因子PU.1在高空ES细胞中的增强表达拯救了表达巨噬细胞限制性标记的破骨细胞和巨噬细胞样吞噬细胞的发育。其他造血系细胞均未生成。它们的发育依赖于M-CSF和RANKL。这些结果表明，PU.1的表达是破骨细胞发生的关键事件，Tal-1在破骨细胞发生过程中可能位于PU.1的上游。较少

项目成果

Presence and distribution of neural crest-derived cells in the murine developing thymus and their potential for differentiation.

小鼠发育胸腺中神经嵴衍生细胞的存在和分布及其分化潜力。

• 发表时间: 2005
• 期刊: Int. Immunol. 17
• 作者: Yamazaki;H.;et al.
• 通讯作者: et al.

Migration of dendritic cells determines divergent immune responses.

树突状细胞的迁移决定了不同的免疫反应。

• 发表时间: 2004
• 期刊: Recent Research Development in Biophysics and Biochemistry (Research Signpost) 4
• 作者: Yoshino;M.;Yamazaki;H.;Hayashi.S.I.
• 通讯作者: Hayashi.S.I.

In vitro differentiation of mouse ES cells into hematopoietic, endothelial, andosteoblastic cell lineages : A possibility of in vitro organogenesis

小鼠 ES 细胞体外分化为造血细胞、内皮细胞、成骨细胞谱系：体外器官发生的可能性

• 发表时间: 2003
• 期刊: Methods Enzymol. 365
• 作者: Tsuneto;M.;et al.
• 通讯作者: et al.

Okuyama, H., et al.: "Discrete types of osteoclast precursors can be generated from embryonic stem cells"Stem Cells. 21. 670-680 (2003)

Okuyama, H.等人：“可以从胚胎干细胞产生离散类型的破骨细胞前体”干细胞。

Tsuneto, M., et al.: "In vitro differentiation of mouse ES cells into hematopoietic, endothelial, and osteoblastic cell lineages : A possibility of in vitro organogenesis"Methods Enzymol.. 365. 98-114 (2003)

Tsuneto, M., 等人：“小鼠 ES 细胞体外分化为造血细胞、内皮细胞和成骨细胞谱系：体外器官发生的可能性”Methods Enzymol.. 365. 98-114 (2003)