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目的体外诱导人外周血单个核细胞成为破骨样细胞。方法从人外周血以密度梯度离心法分离出单个核细胞贴壁培养后分为3组。A组:使用核因子NF-κB受体激活剂配体(RANKL)联合巨噬细胞集落刺激因子(M-CSF)进行诱导;B组:仅使用M-CSF进行诱导;C组:仅使用RANKL进行诱导。14d后使用抗酒石酸酸性磷酸酶(TRAP)染色法以及扫描电镜观察骨片以鉴定细胞是否具有破骨细胞的特征。结果 A组14d时出现较多的多核TRAP染色阳性的细胞,数量为(476±47)个,较B组(54±18)个明显增加(P<0.05),C组不能诱导出TRAP染色阳性细胞。A组诱导生成的细胞具有在骨片上形成骨吸收陷窝的能力,B组骨片上均没有发现明显的骨吸收陷窝。结论从人外周血单个核细胞中可以诱导出破骨样细胞,RANKL和M-CSF是诱导过程中所需要的两个重要因子。
Objective To induce human peripheral blood mononuclear cells to become osteoclast-like cells in vitro. Methods The mononuclear cells isolated from peripheral blood of human by density gradient centrifugation were divided into 3 groups. Group A: induced by nuclear factor NF-κB receptor activator ligand (RANKL) combined with macrophage colony stimulating factor (M-CSF); Group B: induced by M-CSF only; Group C: RANKL Induction. After 14 days, tartrate-resistant acid phosphatase (TRAP) staining and scanning electron microscopy were used to observe whether the cells had osteoclasts. Results The number of multi-nuclear TRAP-positive cells in group A was significantly higher than that in group B (54 ± 18) (P <0.05) on the 14th day in group A (476 ± 47) cell. The cells induced by group A had the ability of forming bone resorption lacuna on the bone slices, and no obvious bone resorption lacuna was found on the bone slices of group B. Conclusion The osteoclast-like cells can be induced from human peripheral blood mononuclear cells, and RANKL and M-CSF are two important factors in the induction process.