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Development of new assay systems for examining the relation between osteoblasts and osteoclasts, and identification of new factors controlling bone metabolism

开发新的检测系统来检查成骨细胞和破骨细胞之间的关系，并鉴定控制骨代谢的新因素

基本信息

批准号：
61870074
负责人：
SUDA Tatsuo
金额：
$ 4.54万
依托单位：
Showa University, School of Dentistry
依托单位国家：
日本
项目类别：
Grant-in-Aid for Developmental Scientific Research
财政年份：
1986
资助国家：
日本
起止时间：
1986 至 1988
项目状态：
已结题

项目摘要

The aim of the present study was to develop new assay systems for examining the relation between osteoblasts and osteoclasts and to identify new factors controlling bone metabolism. In order to address these problems, we performed following experiments: (1) isolation and characterization of osteotropic factors produced by osteoblasts, (2) development of new assay systems for examining osteoclast formation in vitro, and (3) clarification of the role of osteoblasts in osteoclast formation.1. We previously reported that osteoblastic MC3T3-E1 cells produced differentiation inducing factor (DIF) which promoted differentiation of mouse myeloid leukemia cells (M1) into macrophage-like cells. The DIF was partially purified from the conditioned meda of MC3T3-E1 cells and its biological activity in bone resorption was examined. The DIF exhibited a marked bone resorbing activity in a Raisz's assay system and it stimulated osteoclast-like cell formation in a mouse marrow culture system.2. We devel … More oped a mouse bone marrow culture system to examine osteoclast formation in vitro. Bone resorption-stimulating hormones and factors such as 1alpha,25(OH)_2D_3, PTH, PGE_2, IL-1, TNFalpha, and TGF markedly stimulated the formation of osteoclast-like multinucleated cells (MNCs), whereas calcitnin and IFN<@2Y<@D2 strongly inhibited the formation induced by those hormones and factors.3. In order to examine the role of osteoblasts in osteoclast formation, we developed a co-culture system of mouse spleen cells and osteoblastic cells freshly isolated from fetal mouse calvariae. When mouse spleen cells were co-cultured with osteoblastic cells in the presence of 1alpha, 25(OH)_2D_3, osteoclast-like MNCs were formed within 8 days. Neither the same co-culture without the vitamin nor separate cultures of either spleen cells or osteoblastic cells with the vitamin produced osteoclast-like MNCs. These results indicate that osteoblasts are required for differentiation of osteoclast progenitors into multinucleated osteoclasts. Less

本研究的目的是开发新的检测系统，以检查成骨细胞和破骨细胞之间的关系，并确定新的因素控制骨代谢。为了解决这些问题，我们进行了以下实验：（1）分离和鉴定成骨细胞产生的骨促因子，（2）开发新的体外检测破骨细胞形成的检测系统，（3）阐明成骨细胞在破骨细胞形成中的作用。我们曾报道成骨细胞MC 3 T3-E1产生的分化诱导因子（DIF）能促进小鼠髓性白血病细胞（M1）向巨噬细胞样细胞分化。从MC 3 T3-E1细胞的条件梅达中部分纯化DIF，并检测其在骨吸收中的生物学活性。DIF在Raisz's测定系统中表现出显着的骨吸收活性，并刺激小鼠骨髓培养系统中破骨细胞样细胞的形成。2.我们开发了 ...更多信息建立小鼠骨髓细胞体外培养体系，观察破骨细胞的形成。骨吸收刺激激素和因子如1alpha、25（OH）_2D_3、PTH、PGE_2、IL-1、TNF α和TGF显著促进破骨细胞样多核细胞（MNCs）的形成，而降钙素和IFN<@2Y<@D_2则强烈抑制这些激素和因子诱导的MNCs的形成.为了研究成骨细胞在破骨细胞形成中的作用，我们建立了小鼠脾细胞与新鲜分离的胎鼠颅骨成骨细胞共培养体系。当小鼠脾细胞与成骨细胞在1 α，25（OH）_2D_3存在下共培养时，破骨细胞样MNCs在8天内形成。无论是相同的共培养没有维生素，也没有单独的文化，无论是脾细胞或成骨细胞与维生素产生破骨细胞样MNCs。这些结果表明，破骨细胞祖细胞分化成多核破骨细胞所需的成骨细胞。少