高产量高纯度的破骨细胞分离培养  

Isolation of highly enriched and purified rabbit osteoclasts and osteoclast-like cells

作　　者：刘波[1] 于世风[1] 庞淑珍[1] 

机构地区：[1]北京大学口腔医学院病理科,北京100081

出　　处：《中国骨质疏松杂志》2004年第4期409-411,396,共4页Chinese Journal of Osteoporosis

基　　金：国家自然科学基金(39830430)重点项目资助

摘　　要：目的获得高产量高纯度破骨细胞,为骨吸收的体外研究提供丰富的细胞来源。方法 将经典分离乳兔四肢骨破骨细胞的方法与1,25(OH)2D3诱导骨髓干细胞生成破骨样细胞的方法相结合,并应用0.05％胰蛋白酶／0.02％EDTA联合消化的方法将其纯化。结果该方法可诱导生成大量的破骨样细胞,0.05％胰蛋白酶／0.02％EDTA联合消化可使破骨细胞纯化率达95％。诱导生成的破骨样细胞TRAP染色阳性,体外培养具有噬骨能力。结论 该培养方法可产生大量高纯度的破骨样细胞,可为破骨细胞的体外分子生物学研究提供丰富的细胞来源。Objective To obtain highly enriched and purified osteoclasts in enough amount for the study of the mechanism of bone resorption in vitro. Methods The classical osteoclastic isolation method from limbs of rabbits comlnned with the method of induction of the bone marrow cells was used to form osteoclast-like cells in the presence of 1,25(OH)2D3. Mature osteoclasts and osteoclast-like cells were purified by 0.05% trypsin/0.02% EDTA digestion. Results A large number of rabbit osteoclast-like cells and were produced, and 95 % purified osteoclasts were obtained after 0.05% trypsin/0.02% EDTA digestion. The osteoclast-like cells were positive for the TRAP staining and capable of bone resorption in vitro. Conclusions This method can produce highly enriched and purified osteoclasts in enough amount for biochemical and molecular biological research.

关 键 词：破骨细胞 破骨样细胞 诱导 细胞来源 联合 分离培养 四肢骨 胰蛋白酶 体外研究 骨吸收 

分 类 号：R681[医药卫生—骨科学] R783.5[医药卫生—外科学]