目的 用核因子-κB受体活化因子配体(RANKL)诱导破骨细胞前体细胞分化成熟,建立获取成熟破骨细胞的方法.方法 用破骨细胞前体细胞RAW264.7细胞为模型,RANKL诱导培养4～9 d,抗酒石酸酸性磷酸酶(TRAP)染色观察TRAP阳性多核细胞形成,罗丹明-鬼笔环肽荧光染色观察纤维性肌动蛋白(F-actin)环,DAPI染色观察细胞核,甲苯胺蓝染色观察牛骨片表面的吸收陷窝情况.结果 RANKL可诱导RAW264.7细胞形成TRAP染色阳性的多核细胞,形成F-actin环,骨片吸收陷窝明显.结论 RANKL可诱导RAW264.7细胞向成熟破骨细胞分化,该诱导模型可用于破骨细胞分化研究.%Objective To explore the method of getting mature osteoclasts by RANKL inducing osteoclast differentiation and maturity in RAW264. 7 cells. Methods RAW264. 7 cells were treated with RANKL for 4 or 9 days, TRAP-positive multinuclear cells were examined by TRAP staining and F-actin rings were observed by fluorescent microscope with rhodamine-Phalloidin. Osteoclastic absorption function was determined by bone resorption pits formation on calf cortex slice with toluidine blue staining. Results RAW264. 7 cells could be were induced to be TRAP-positive multinuclear cells, and to form F-actin rings. Resorption pits made by cells treated with RANKL were more than that of without RANKL significantly. Conclusions RANKL could induce osteoclast differentiation and maturity in RAW264. 7 cells. And it could be used as a models for research of osteoclast differentiation research.
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