Udagawa N, Takahashi N, Akatsu T, Tanaka H, Sasaki T, Nishihara T, Koga T, Martin T J, Suda T
Department of Biochemistry, School of Dentistry, Showa University, Tokyo, Japan.
Proc Natl Acad Sci U S A. 1990 Sep;87(18):7260-4. doi: 10.1073/pnas.87.18.7260.
We previously reported that osteoclast-like cells were formed in cocultures of a mouse marrow-derived stromal cell line (ST2) with mouse spleen cells in the presence of 1 alpha, 25-dihydroxyvitamin D3 and dexamethasone. In this study, we developed a new coculture system to determine the origin of osteoclasts. When relatively small numbers of mononuclear cells (10(3)-10(5) cells per well) obtained from mouse bone marrow, spleen, thymus, or peripheral blood were cultured for 12 days on the ST2 cell layers, they formed colonies with a linear relationship between the number of colonies formed and the number of hemopoietic cells inoculated. Tartrate-resistant acid phosphatase (TRAPase)-positive mononuclear and multinucleated cells appeared in the colonies (TRAPase-positive colonies) in response to 1 alpha, 25-dihydroxyvitamin D3 and dexamethasone. When hemopoietic cells suspended in a collagen-gel solution were cultured on the ST2 cell layers to prevent their movement, TRAPase-positive colonies were similarly formed, indicating that each colony originated from a single cell. All of the colonies consisted of nonspecific esterase-positive cells. The monocyte-depleted population prepared from peripheral blood failed to form colonies, whereas the monocyte-enriched population produced a large number of TRAPase-positive colonies. In addition, alveolar macrophages formed TRAPase-positive colonies most efficiently on the ST2 cell layers in the presence of the two hormones. Salmon 125I-labeled calcitonin specifically bound to the TRAPase-positive cells. Resorption lacunae were formed on dentine slices on which cocultures were performed. When direct contact between the peripheral blood cells and the ST2 cells was inhibited by a collagen-gel sheet, no TRAPase-positive cells were formed. These results indicate that osteoclasts are also derived from the mature monocytes and macrophages when a suitable microenvironment is provided by bone marrow-derived stromal cells.
我们先前报道，在1α,25 - 二羟基维生素D3和地塞米松存在的情况下，小鼠骨髓来源的基质细胞系（ST2）与小鼠脾细胞共培养时可形成破骨细胞样细胞。在本研究中，我们开发了一种新的共培养系统以确定破骨细胞的起源。当将从小鼠骨髓、脾脏、胸腺或外周血中获取的相对少量的单核细胞（每孔10³ - 10⁵个细胞）在ST2细胞层上培养12天时，它们形成集落，形成的集落数量与接种的造血细胞数量呈线性关系。在1α,25 - 二羟基维生素D3和地塞米松作用下，集落中出现抗酒石酸酸性磷酸酶（TRAPase）阳性的单核和多核细胞（TRAPase阳性集落）。当将悬浮于胶原凝胶溶液中的造血细胞在ST2细胞层上培养以防止其移动时，同样形成了TRAPase阳性集落，表明每个集落起源于单个细胞。所有集落均由非特异性酯酶阳性细胞组成。从外周血制备的单核细胞耗竭群体未能形成集落，而单核细胞富集群体产生大量TRAPase阳性集落。此外，在两种激素存在的情况下，肺泡巨噬细胞在ST2细胞层上最有效地形成TRAPase阳性集落。鲑鱼¹²⁵I标记的降钙素特异性结合TRAPase阳性细胞。在进行共培养的牙本质切片上形成了吸收陷窝。当用胶原凝胶片抑制外周血细胞与ST2细胞之间的直接接触时，未形成TRAPase阳性细胞。这些结果表明，当骨髓来源的基质细胞提供合适的微环境时，破骨细胞也来源于成熟的单核细胞和巨噬细胞。
