小鼠破骨细胞样细胞的诱导培养及重组ODF胞外片段对其影响  

Cultivation of Osteoclast-like Cell and Effect of Ectodomain of Recombinant Murine ODF on Osteoclast-like Cell Formation

作　　者：戴芳[1] 王宝利[1] 梁晖[1] 戴晨琳[1] 邱明才[1] 

机构地区：[1]天津医科大学总医院,天津医科大学内分泌所,300052

出　　处：《天津医药》2004年第10期630-632,共3页Tianjin Medical Journal

基　　金：天津市自然科学基金资助项目 (项目编号 :043606811);天津市高等学校科技发展基金项目 (项目编号 :020108)

摘　　要：目的 :建立破骨细胞样细胞 (OLC)的体外培养方法并观察本室制备的重组小鼠破骨细胞分化因子(ODF)胞外片段对OLC生成的影响。方法:在无菌条件下取小鼠股骨 ,收集骨髓细胞 ,在重组小鼠ODF胞外片段和单核 -巨噬细胞集落刺激因子 (M -CSF)作用下诱导骨髓细胞向OLC分化。37℃5 %CO2 培养7d后行抗酒石酸酸性磷酸酶 (TRAP)染色 ,光镜下观察OLC生成情况 ,同时行扫描电镜鉴定。结果:ODF胞外片段诱导培养体系中有多核的OLC生成 ,且呈剂量依赖性 ,多组比较差别有统计学意义 (P<0.05)。结论:本实验建立的小鼠髓源OLC体外诱导培养方法 ,证实重组小鼠ODF胞外片段可显著促进OLC的生成。Objective:To establish a method for murine osteoclast-like cells(OLC)cultivation in vitro and observe the effect of ectodomain of recombinant murine osteoclast differentiation factor(rmODF)on OLC for-mation.Methods:Experimental mice were sacrificed and the femora were removed from these mice aseptically.Both the two epiphyseal ends of the femora were cut off,and the marrow cavities were flushed with D-Hanks.The collected bone marrow cells were washed twice with D-Hanks and cultured in a-MEM containing10%FBS(fetal bovine serum),80ng/mL M-CSF and divert concentration of ectodomain of rmODF in a96-well plate(37℃,5%CO 2 )for7days.Bone slices were embedded into the cultured plate before the bone marrow cells were added.The cultured cells were stained for tartrate-resistant acid phosphatase(TRAP).TRAP-pos-itive multinucleated cells were observed under light microscopy and counted.Meanwhile,scanning electron microscopy was used to identify OLCs and the pits of bone resorption.Results:In the presence of M-CSF and ectodomain rmODF,OLCs were present in the cultured plate based on the observation of cell morphology,the staining for TRAP and the pits of bone resorption.The number of the ODF-induced OLCs increased in a dose-dependant manner(P<0.05).Conclusion:The method of cultivating marrow-derived osteoclast-like cell in vitro was established.The ectodomain of rmODF could induce the formation of OLC significantly.

关 键 词：小鼠 破骨细胞样细胞 OLC 骨髓细胞 M-CSF 抗酒石酸酸性磷酸酶 诱导培养 培养方法 结论 情况 

分 类 号：R783.5[医药卫生—口腔医学] R361.3[医药卫生—临床医学]