成人破骨细胞的体外培养  

Culture of human osteoclasts

作　　者：欧阳桂林[1] 杨庆铭[1] 邓廉夫[1] 许福萍[1] 朱雅萍[1] 

机构地区：[1]上海第二医科大学附属瑞金医院骨科,上海市伤骨科研究所上海200025

出　　处：《中西医结合学报》2003年第1期57-61,共5页Journal of Chinese Integrative Medicine

基　　金：上海市科学技术委员会科技攻关项目 (964 1190 2 4)

摘　　要：目的　建立成人破骨细胞体外培养的方法 ,观察成骨细胞对破骨细胞分化、增殖和功能的影响 ,为进一步研究补肾中药防治骨溶解和骨质疏松症提供基础。材料和方法　从成人骨髓中提取单核细胞 ,以 1,2 5 (OH) 2 D3 作为刺激因子 ,分 3组培养 :单核细胞与成骨细胞共同培养 ,单核细胞单独培养 ,单核细胞与成骨细胞条件培养液培养。生物显微镜下观察细胞的分化过程 ,HE染色和抗酒石酸酸性磷酸酶 (Trap)染色方法作组织化学观察 ,扫描电镜 (SEM)观察骨吸收功能。 结果　在单核细胞与成骨细胞共同培养组 ,可见单核细胞逐步融合成多核细胞 ;HE染色和Trap染色可见阳性多核细胞 ;扫描电镜观察可见骨片上有骨吸收陷窝形成。其他两组则未得到上述结果。结论　成人骨髓单核细胞在一定培养条件下能分化成熟为具有典型的破骨细胞表型的多核细胞 :Trap染色阳性 ,具有骨吸收功能。与成骨细胞的直接接触是破骨细胞分化、增殖和发挥功能的必不可少的条件。Objective To establish a stable, useful culture system for human osteoclasts and to investigate the effect of osteoblasts on the differentiation, proliferation and activation of osteoclasts so as to provide a base for the studies on prevention and treatment of osteolysis and osteoporosis. Methods In the presence of 1,25 (OH) 2D 3, monocytes abstracted from human bone marrow were cultured in three groups:co culture of monocytes and osteoblasts,monocytes alone,monocytes with conditional media(CM) of osteoblasts.Differentiation process of the cultured cells was observed under biological microscope.HE staining and tartrate resistant acid phosphatase (Trap) staining were employed to assay the cultured cells.The resorption pits on bone slices,on which cells were cultured, were observed under scanning electronic microscope (SEM). Results In group of co culture of monocytes and osteoblasts, monocytes gradually fused to form multinucleated cells (MNCs),and the MNCs were also indicated in HE staining and Trap staining.The SEM showed a number of resorption pits on bone slices .In the other two groups,Trap positive MNCs were not obtaind,and resorption pits were not observed on bone slices. Conclusion In this culture, monocytes obtained from human marrow fused to form multinucleated cells(MNCs) that express the main characteristics of the osteoclast phenotype,such as Trap positive and the ability to form resorption lacunae when cultured on bone slices.Cell to cell contact with osteoblasts was necessary for the differentiation,proliferation and activation of osteoclasts.

关 键 词：破骨细胞 体外培养 细胞培养 成骨细胞 骨溶解 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]