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）破骨细胞生成对骨髓形成的分析：破骨细胞是造血细胞，参与骨吸收和骨重塑。 NF-κB配体受体激活剂(RANKL)和巨噬细胞集落刺激因子(M-CSF)对于破骨细胞的发育至关重要。 Toll 样受体 (TLR) 家族与 RANK 共享一些下游信号传导，然而，从未有报道称通过 TLR 的信号传导在没有 RANKL 的情况下诱导破骨细胞生成。我们发现，在存在 M-CSF 和 LPS（一种 TLR4 配体）且不添加 RANKL 的培养物中，蛋白酪氨酸磷酸酶 SHP-1 缺陷的 me^v/me^v 骨髓细胞产生了大量成熟的破骨细胞。破骨细胞前体存在于 BM 细胞的 Kit 阳性细胞富集部分中。尽管 me^v/me^v 骨髓细胞需要与野生型细胞相当浓度的 RANKL 或 TNF-α 来启动破骨细胞生成，但在 M-CSF 存在的情况下，me^v/ … 更多 me^v 骨髓培养物中的多核破骨细胞数量显着增加。这些结果表明，SHP-1功能缺陷不仅会加速RANKL/RANK的生理性破骨细胞发育，而且还会获得LPS破骨细胞生成的异常途径。2)诱导无血胚胎干(ES)细胞系造血：转录因子Tal-1对于造血发育的规范至关重要。缺乏 Tall 的小鼠无法产生任何造血前体细胞。使用我们的基质细胞共培养系统，我们证明在 Tall-null ES 细胞中四环素控制下转录因子 PU.1 的强制表达可以挽救表达巨噬细胞限制标记的破骨细胞和巨噬细胞样吞噬细胞的发育。没有产生其他造血谱系细胞。它们的发育依赖于 M-CSF 和 RANKL。这些结果表明 PU.1 的表达是破骨细胞生成的关键事件，并且 Tal-1 在破骨细胞生成过程中的调控层次中可能位于 PU.1 的上游。较少的

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)