Extablishment of osteoclast cell line and cytological and immunological anaylysis on differentiation of pre-osteoclast.

破骨细胞系的建立及前破骨细胞分化的细胞学和免疫学分析。

• 批准号: 04454464
• 负责人: SUZUKI Akitoshi
• 金额: $ 4.35万
• 依托单位: Niigata University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (B)
• 财政年份: 1992
• 资助国家: 日本
• 起止时间: 1992 至 1993
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-04454464/
• 关键词: 1,25-Dihydroxyvitamin D_3; Colony-stimulating factor; Macrophage, Granulocyte differentiation; 前破骨細胞様ハイブリドーマ; 活性型ビタミンD_3; M-CSF; GM-CSF; マクロファージ; 顆粒球

1,25-Dihydroxyvitamin D_3 (1,25(OH)_2D_3) was recently shown to promote maturation fo 5-fluorouracil (5FU)-treated bone marrow cells by up-regulating macrophage-clony stimulating factor (M-CSF) receptors in the presence of interleukin 1alphsa (IL-1alpha). In order to reveal how1,25(OH)_2D_3 interacts with clony-stimulation factors and regulates the differentiation of bone marrow progenitor cell population, in the present study, nutural bone marrow cells were isolated from untreated mice and used in alpha-minimum essential medilum supplelmented with 20% heat-inactivated horse serum without added appropriate cytokines. Under the conditions, cells spontaneously differentiated gradually with days of culture, as assessed by epression of macrophage differentiation antigens such as Mac-1 (CD11b) and F4/80. Both M-CSF and granulocyte macrophage-colony stimulating factor (GM-CSF) induced only Mac-1 antigen expression. Simultaneous treatment with M-CSF and 1,25(HD)_2D_3 per se has no effect on the espression for up to 11 days. In addition, successive treatment with 1,25(OH)_2D_3 and M-CSF or GM-CSF dramatically enhanced expression of both antigens or Mac-1 antigen, respectively. Similarly, both simultaneous and successive treatment with 1,25(HD)_2D_3 and M-CSF significantly enhanced phagocytic activity and H_2O_2 production, Whereas successive treatment with 1,25(HD)_2D_3 and GM-CSF significantly enhanced only phagocytic activity. Enzymehistochemical study demonstrated that cells treated simultaneously or successively with 1,25(HD)_2D_3 and M-CSF were strongly positive for nonspecific esterase (NSE), a macrophage-specific maker, and that simultaneous or succesive treatment with 1,25(HD)_2D_3 primes bone marrow progenitor cell populations not only to M-CSF but also to GM-CSF and thereby accelelrates the CSFs-dependent differentiation of the cell

1,25-二羟基维生素D_3 （1,25(OH)_2D_3）最近被证明在白细胞介素1 α (il -1 α)存在下，通过上调巨噬细胞克隆刺激因子（M-CSF）受体，促进5-氟尿嘧啶（5FU）处理的骨髓细胞的成熟。为了揭示1,25(OH)_2D_3如何与克隆刺激因子相互作用并调节骨髓祖细胞群的分化，本研究从未经处理的小鼠中分离出天然骨髓细胞，并将其用于添加20%热灭活马血清的α -minimum必需培养基中，不添加适当的细胞因子。在此条件下，通过巨噬细胞分化抗原Mac-1 （CD11b）和F4/80的表达，细胞随培养时间逐渐自发分化。M-CSF和粒细胞巨噬细胞集落刺激因子（GM-CSF）均仅诱导Mac-1抗原表达。同时用M-CSF和1,25(HD)_2D_3本身治疗，在长达11天的时间内对表达没有影响。此外，1,25(OH)_2D_3和M-CSF或GM-CSF的连续处理分别显著提高了抗原或Mac-1抗原的表达。同样，125 (HD)_2D_3和M-CSF同时和连续处理均显著提高了吞噬活性和H_2O_2的产生，而125 (HD)_2D_3和GM-CSF连续处理仅显著提高了吞噬活性。酶组织化学研究表明，同时或先后用1,25(HD)_2D_3和M-CSF处理的细胞非特异性酯酶（NSE）呈强烈阳性，这是一种巨噬细胞特异性制造物，同时或连续用1,25(HD)_2D_3处理的骨髓祖细胞群不仅对M-CSF也对GM-CSF，从而加速了细胞对csf的依赖性分化

项目成果

Orikasa, M.et al.: "Induction of macrophagic and granulocytic differentiation of murine bone marrow progenitor cells by 1,25-Dihydroxyvitamin D_3" Calcif.Tissue Int.53. 193-200 (1993)

Orikasa, M.等人：“1,25-二羟基维生素 D_3 诱导小鼠骨髓祖细胞的巨噬细胞和粒细胞分化”Calcif.Tissue Int.53。

Orikasa,M.et al.: "Induction of macrophagic and granulocytic differentiation of murine bone marrow progenitor cells by 1,25-dihydroxyvitamin D_3." Calcified Tissue International. 53. 193-200 (1993)

Orikasa,M.et al.：“1,25-二羟基维生素 D_3 诱导小鼠骨髓祖细胞的巨噬细胞和粒细胞分化。”