Kukita A, Kukita T, Hata K, Kurisu K, Kohashi O
Department of Microbiology, Saga Medical School, Japan.
Bone Miner. 1993 Nov;23(2):113-27. doi: 10.1016/s0169-6009(08)80048-7.
To study the effects of osteoblast products on osteoclast formation, we added the conditioned medium (CM) of rat osteoblastic cell line ROS17/2.8 to rat bone marrow cultures, in which tartrate-resistant acid phosphatase (TRAP)-positive osteoclast-like multinucleate cells (MNCs) formed in the presence of 10(-8) M 1,25-dihydroxyvitamin D3 (1,25(OH)2D3). The formation of 1,25(OH)2D3-dependent TRAP-positive MNC at day 7 of culture was strongly inhibited by the > 10 kDa fraction of ROS17/2.8 cell-CM (ROSCM), but heat treated ROSCM (htROSCM) expressed marked stimulation in the formation of the MNCs. The expression of several osteoclastic phenotypes of the MNCs induced by htROSCM and 1,25(OH)2D3 was more enhanced compared with that of the MNCs induced by 1,25(OH)2D3 alone. The MNCs induced by htROSCM and 1,25(OH)2D3 were highly motile, were sensitive to calcitonin (CT), and had high bone resorbing activity. These data suggest that htROSCM promotes the osteoclast differentiation in the presence of 1,25(OH)2D3 in a rat bone marrow culture system. The stimulatory activity of TRAP-positive MNC formation in htROSCM is derived from heat-stable protein(s) that is (are) thought to be different from colony-stimulating factors (CSFs) such as macrophage-CSF (M-CSF) or granulocyte-macrophage-CSF (GM-CSF).
为研究成骨细胞产物对破骨细胞形成的影响，我们将大鼠成骨细胞系ROS17/2.8的条件培养基（CM）添加到大鼠骨髓培养物中，在含有10(-8) M 1,25 - 二羟基维生素D3（1,25(OH)2D3）的情况下，可形成抗酒石酸酸性磷酸酶（TRAP）阳性的破骨细胞样多核细胞（MNCs）。培养第7天时，ROS17/2.8细胞 - CM（ROSCM）分子量> 10 kDa的组分强烈抑制了1,25(OH)2D3依赖性TRAP阳性MNC的形成，但热处理的ROSCM（htROSCM）对MNCs的形成有明显的刺激作用。与单独由1,25(OH)2D3诱导的MNCs相比，htROSCM和1,25(OH)2D3诱导的MNCs的几种破骨细胞表型的表达增强更为明显。htROSCM和1,25(OH)2D3诱导的MNCs具有高迁移性，对降钙素（CT）敏感，并且具有高骨吸收活性。这些数据表明，在大鼠骨髓培养系统中，htROSCM在1,25(OH)2D3存在的情况下促进破骨细胞分化。htROSCM中TRAP阳性MNC形成的刺激活性源自热稳定蛋白，这些蛋白被认为不同于集落刺激因子（CSF），如巨噬细胞集落刺激因子（M - CSF）或粒细胞 - 巨噬细胞集落刺激因子（GM - CSF）。
