目的:从人外周血单个核细胞诱导培养获得高产量高纯度破骨细胞,为破骨细胞的体外研究提供丰富的细胞来源。方法:从外周血分离单个核细胞贴壁培养,采用0.25%胰蛋白酶/0.02%EDTA联合消化,纯化后以RANKL和M-CSF加以诱导(A组),并与未经消化之传统方法(B组)比较。结果:与B组相比,A组可获得(1426±204)个破骨细胞(P<0.05),0.25%胰蛋白酶/0.02%EDTA联合消化可使破骨细胞纯化率达90%。诱导生成的破骨细胞TRAP染色阳性,功能试验显示具有噬骨能力。结论:联合消化结合RANKL/M-CSF诱导培养法可产生大量的破骨样细胞,方法简便而且经济实用。 OBJECTIVE: To obtain high-yield and high-purity osteoclasts induced from human peripheral blood mononuclear cells and to provide a rich source of cells for in vitro studies of osteoclasts. Methods: Mononuclear cells isolated from peripheral blood were adherently cultured, digested with 0.25% trypsin / 0.02% EDTA, purified and induced with RANKL and M-CSF (group A), and compared with the conventional method without digestion Group) comparison. Results: Compared with group B, (1426 ± 204) osteoclasts were obtained in group A (P <0.05). Combined with 0.25% trypsin / 0.02% EDTA, the osteoclasts could be purified by 90%. Induced osteoclast TRAP positive staining, functional tests showed that the ability to bite bone. Conclusion: Combined digestion and RANKL / M-CSF induction culture method can produce a large number of osteoclast-like cells. The method is simple and economical.