人破骨细胞的体外培养及鉴定  

Culture and identification of human osteoclasts in vitro

作　　者：刘水涛[1] 雷鸣[2] 安佰京[1] 杨军[1] 张鹏礼[1] 邢更彦[1] 

机构地区：[1]武警总医院骨科,北京100039 [2]河北大学附属医院,石家庄071000

出　　处：《武警医学》2011年第5期381-384,共4页Medical Journal of the Chinese People's Armed Police Force

基　　金：武警总医院Ⅰ类课题(WZ 2009052)

摘　　要：目的探索人破骨细胞的体外培养及鉴定方法 ,为各类试验研究提供大量高活性破骨细胞。方法分离骨髓血得到单个核细胞,用不同培养液、诱导剂在不同浓度下培养破骨细胞,通过TRAP染色鉴定破骨细胞形态,ELISA法检测特异性蛋白表达水平,扫描电镜分析骨片骨陷窝面积。结果用α-MEM培养液,10^(-7)mmol/L浓度的1,25(OH)_2VD_3或50ng/ml浓度的RANKL和30ng/ml的M-CSF作为诱导剂,所培养的破骨细胞多核、特异性蛋白表达水平高、蚀骨能力强。结论 用DMEM培养液和α-MEM培养液培养均是一种能在体外诱导生成大量人破骨细胞的方法。同时,建议采用1,25(OH)_2VD_3作为诱导剂。Objective To establish an in vitro method of culture and identification of human osteoclasts, and to provide a large number of highly - active osteoclasts for experimental researches. Methods Mononuclear cells were obtained from bone marrow. The osteoclasts were cultured in different media, by different inducers and at different concentrations. Morphological characteristics of cells were identified by TRAP staining, and specific proteins by ELISA. The ability of resorption was analysed by scanning electron microscopy. Results In group C and D, the α -MEM was used as culture medium and 1, 25（OH）2VD3（10^-7 mmol/L） or RANKL （50 ng/ml） and M- CSF （30 ng/ml） as inducers. The osteoclasts were multinuclear and highly -active. Conculsions A method has been found to cultivate numerous highly - active human osteoclasts in vitro.

关 键 词：人破骨细胞 细胞培养 TRAP染色 

分 类 号：R494.26[医药卫生—康复医学]