中文摘要：

目的用胰酶消化法纯化获取大量高纯度破骨细胞，为进行相关分子生物学研究奠定基础。方法以1,25-（OH）2D3/地塞米松诱导培养不同年龄段（1、12、24、36d）SD大鼠的骨髓破骨细胞，采用0．25％胰蛋白酶，0．02％乙二胺四乙酸纯化。通过倒置相差显微镜观察细胞形态，采用抗酒石酸酸性磷酸酶（Trap）染色和扫描电镜观察鉴定破骨细胞，并计数分析。结果1、12、24、36d组大鼠骨髓破骨细胞均培养成功，经Trap染色和扫描电镜观察证实为体外有噬骨能力的破骨细胞，纯化率达到90％。1d和12d组破骨细胞出现较早且数量较多，细胞计数两组间无统计学差异（P〉0．05），但它们与其余各组均有统计学差异（P〈0．05）。结论采用胰酶消化法纯化1,25-（0H）2D3/地塞米松诱导培养的大鼠骨髓破骨细胞，纯度较高；选用大鼠以10-12d龄较为适宜。

英文摘要：

Objective To obtain highly enriched osteoclasts in vitro. Methods The bone marrow ceils of 1-day, 12-day, 24-day and 36-day old Sprague-DawleyCSD） rats were separately cultured with the presence of 1,25-（OH）2D3 and dexamethasone, and the osteoclast-like cells were identified by Trap staining and scanning electron microscope observation and were purified by trypsin digestion. The count of Trap positive osteoclast-like cells was analyzed statistically. Results The osteoclast-like cells of each group were Trap positive cells and could form the bone absorption lacunas in vitro. The amounts of osteoclast-like cells were different statistically between the groups （P〈0.05） except between the groups of 1-day old and 12-day old（P〈0.05）. Highly enriched osteoclast-like cells were harvested by the digestion of 0.25% trypsin and 0.02% EDTA. Conclusion It＇s indicated that a large amount of highly enriched osteoclast-like cells could be obtained through the culture of bone marrow cells of 10-day and 12-day old SD rats with 1,25- （OH） 2D3 and dexamethasone and the digestion of trypsin/EDTA.