Takahashi N, Yamana H, Yoshiki S, Roodman G D, Mundy G R, Jones S J, Boyde A, Suda T
Department of Biochemistry and Oral Pathology, School of Dentistry, Showa University, Tokyo, Japan.
Endocrinology. 1988 Apr;122(4):1373-82. doi: 10.1210/endo-122-4-1373.
We developed a mouse bone marrow culture system to examine the process of osteoclast-like multinucleated cell formation from its progenitors. When mouse marrow cells were cultured for 8 days with 1 alpha,25-dihydroxyvitamin D3 [1 alpha,25-(OH)2D3, 10(-10) to 10(-7) M] or human PTH (1-34) (25-100 ng/ml), tartrate-resistant acid phosphatase (TRACP)-positive multinucleated cells formed. No TRACP-positive multinucleated cells appeared in the absence of these hormones. 1 alpha,25-(OH)2D3 and PTH also increased the number of the clusters of TRACP-positive mononuclear cells. Time course studies showed that these TRACP-positive mononuclear cell clusters appeared before the formation of TRACP-positive multinucleated cells, suggesting that the TRACP-positive mononuclear cells are precursors of the multinucleated cells. Salmon calcitonin markedly inhibited the formation of TRACP-positive multinucleated cells but not TRACP-positive mononuclear cell clusters induced by 1 alpha,25-(OH)2D3 or PTH. TRACP-positive mononuclear cells and multinucleated cells were rarely stained for nonspecific esterase, but some mononuclear cells were positively stained for both nonspecific esterase and TRACP. More that 90% of the TRACP-positive mononuclear cell clusters and multinucleated cells were found near colonies of alkaline phosphatase-positive mononuclear cells (possibly osteoblasts). When marrow mononuclear cells were cultured on sperm whale dentine slices in the presence of 1 alpha,25-(OH)2D3 or PTH, numerous resorption lacunae were formed. These results suggest that 1) TRACP-positive multinucleated cells formed in response to osteotropic hormones in mouse marrow cultures satisfy most of the criteria of osteoclasts, and 2) osteoblasts may play an important role in osteoclast formation.
我们开发了一种小鼠骨髓培养系统，以研究破骨细胞样多核细胞从其祖细胞形成的过程。当小鼠骨髓细胞与1α,25 - 二羟基维生素D3 [1α,25-(OH)2D3，10(-10)至10(-7)M]或人甲状旁腺激素（1 - 34）（25 - 100 ng/ml）一起培养8天时，形成了抗酒石酸酸性磷酸酶（TRACP）阳性的多核细胞。在没有这些激素的情况下，未出现TRACP阳性的多核细胞。1α,25-(OH)2D3和甲状旁腺激素也增加了TRACP阳性单核细胞簇的数量。时间进程研究表明，这些TRACP阳性单核细胞簇在TRACP阳性多核细胞形成之前出现，这表明TRACP阳性单核细胞是多核细胞的前体。鲑鱼降钙素显著抑制由1α,25-(OH)2D3或甲状旁腺激素诱导的TRACP阳性多核细胞的形成，但不抑制TRACP阳性单核细胞簇的形成。TRACP阳性单核细胞和多核细胞很少被非特异性酯酶染色，但一些单核细胞同时被非特异性酯酶和TRACP阳性染色。超过90%的TRACP阳性单核细胞簇和多核细胞在碱性磷酸酶阳性单核细胞（可能是成骨细胞）的集落附近被发现。当骨髓单核细胞在1α,25-(OH)2D3或甲状旁腺激素存在的情况下在抹香鲸牙本质切片上培养时，形成了大量的吸收陷窝。这些结果表明：1）在小鼠骨髓培养物中，响应促骨激素形成的TRACP阳性多核细胞满足破骨细胞的大多数标准；2）成骨细胞可能在破骨细胞形成中起重要作用。
