小鼠破骨细胞骨髓诱导模型的建立及功能观察

Establishment of experimental model of osteoclast differentiation induced by bone marrow mononuclear phagocytes of mouse

目的获得大量高质量小鼠破骨细胞,为体外研究破骨细胞骨吸收功能提供丰富的细胞来源。方法采用巨噬细胞集落刺激因子(macrophage colony stimulating factor,M-CSF)和破骨细胞分化因子(receptor activator of nuclear factor-κB ligand,RANKL)诱导小鼠骨髓单核细胞分化为破骨细胞,并通过抗酒石酸酸性磷酸酶(tartrate-resistant acid phos-phatase,TRAP)染色和骨吸收实验来鉴定破骨细胞及其噬骨能力。结果诱导3d后可见TRAP(+)多核细胞出现,诱导5d后骨片上可见蓝紫色的吸收陷窝。随着培养时间的延长,TRAP(+)多核细胞数目和吸收陷窝呈现时间依赖性增长趋势(P<0·05)。结论M-CSF和RANKL诱导小鼠骨髓单核细胞可产生大量的具有活跃噬骨能力的破骨细胞。

Objective To obtain plenty of pure osteoclasts for researching the function of osteoclasts in vitro. Methods Osteoclasts were induced by bone marrow mononuclear phagocytes of mouse using medium with macrophage colony stimulating factor （M-CSF） and receptor activator of nuclear factor-kappaB ligand （RANKL）. The tartrate resistant acid phosphatase （TRAP） staining and bone resorption experiment were used to identify the osteoclasts. Results The TRAP positive multinuclear ceils were induced from mouse bone marrow mononuclear phagocytes after 3 days of inoculation, and plenty of resorption pits formed on bone slices after 7 days of inoculation. The number of TRAP positive multinuclear ceils and bone resorption pits gradually increased during the period of induction. Conclusion The osteoclasts were successfully induced from bone marrow mononuclear phagocytes in the presence of M-CSF and RANKL.