人成骨细胞与外周血单核细胞共同培养诱导破骨细胞样细胞

Human primary osteoblasts and peripheral blood monocytes coculture to induce osteoclast-like cells

目的 探讨人外周血单核细胞与原代人成骨细胞共同培养体外转化为破骨细胞样细胞的条件 ,并观察其体外生长、功能情况。方法 分别分离培养人成骨细胞 ,外周血单核细胞 ,并在含1,2 5 (OH) 2 D3(10 -7mol/L) ,地塞米松 (10 -8mol/L)及巨噬细胞集落刺激因子 (M CSF) (2 5ng/ml)的条件液中共同培养 ,用相差显微镜 ,抗酒石酸酸性磷酸酶染色 (TRAP)观察破骨细胞样细胞生长情况 ,应用扫描电镜 (SEM)观察骨片陷窝以反映其功能。结果 共同培养中的单核细胞逐渐融合 ;TRAP染色阳性多核细胞在第 7天明显增多 ;骨片陷窝呈现多种形态。结论 人原代成骨细胞与人外周血单核细胞共同培养可以诱导出破骨细胞样细胞 。

Objective To explore the culture condition for coculture of primary human osteoblasts(OBs)and monocytes(MCs)in blood to induce osteoclast like cells and observe their growth and function Method OBs and MCs isolated separately were cocultured in conditioned medium containing 1,25(OH) 2D 3(10 -7 mol/L),dexamathasone(10 -8 mol/L) and MCSF(25 ng/ml) The growth and function of osteoclast like cells were assessed by phase contrast microscope,TRAP staining and SEM Results MCs fused to form multinucleated cells around OBs Many TRAP positive multinucleated cells appeared after 7 days The pits on bone slices showed various forms Conclusion The coculture of primary OBs and MCs in blood can induce osteoclast like cells,while the differentiation stage of OBs and plating density are vital points for good results