目的研究人外周血单个核细胞体外诱导成为破骨细胞样细胞阳性率。方法将人外周血以密度梯度离心方法分离出单个核细胞,实验组在地塞米松、1,25(OH)2D3和M—CSF的存在下,在盖玻片和骨磨片上与UMR106细胞共同培养20天,对照组除不与UMR106共培养外,其余条件与实验组相同。结果在培养20天后,实验组电镜检查发现有多核巨噬细胞形成,有伪足,骨磨片上有骨吸收陷窝形成,盖玻片上有TRAP染色阳性细胞形成,其细胞阳性率约为3%,对照组没有TRAP阳性细胞形成。结论人外周血单个核细胞在地塞米松、1,25(OH)2D3和M-CSF的存在下,与UMR106细胞共同培养有TRAP阳性细胞形成,其细胞阳性率约为3%,提示可能在单核,巨噬细胞分化较早阶段就决定了哪些细胞能被诱导成为OLC。
Objective To discuss the meaning of tartrate resistant acid phosphatase (TRAP) positive cells ratio while human peripheral blood cells are induced to osteoclast-like cells in vitro .Method Mononucleated cells were isolated by means of density gradient centrifugation from 20ml peripheral blood collected from healthy normal donors. In the study group, The mononucleated cells were planted on the glass coverships or bone slices and cocuhured with UMR 106 cell line, DMEM which contented 1, 25 dihydroxyvitamin D3, M-CSF and dexamethasone was used as the culture medium. The control group used the same culture medium but did not cocuhure with UMR 106.Result After 20 days of culture, the polynuclear macrophage with pseudopodium were found under electric microscope scan in study group, and bone absorb pits were found in the bone slice also. 6 TRAP-positive cells were found on the slices per 200 mononucleate in staining. In control group, we did not find the same result. Conclusion Our study showed that about 3 % mononucleated cells of human peripheral blood could be induced to osteoclast-like cells which have the ability of bone resorption and show TRAP-positive when cocuhured with UMR 106 in the presence of 1,25 dihydroxyvitamin D3, macrophage-colony stimulating factor( MCSF) and dexamethasone, suggesting that which kind of cells could be induced to OLC is decided in their early stage.