RANKL诱导破骨细胞前体细胞分化成熟

RANKL inducing osteoclast differentiation and maturity in RAW264.7 cells

目的用核因子-KB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4～9d，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-aetin rings. Resorption pits made by ceils 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 osteoelast differentiation research.