人骨肉瘤原代细胞条件培养基中骨形态蛋白(BMP)的实验研究  

Experimental Study of Bone Morphogenetic Protein (BMP) in Conditioned Medium of Human Osteosarcoma Cell

作　　者：冷燕奎[1] 吴华[1] 陈安民[1] 瞿智玲[2] 陈继革[1] 

机构地区：[1]华中科技大学同济医学院附属同济医院骨科,武汉430030 [2]华中科技大学同济医学院病理系

出　　处：《中国矫形外科杂志》2004年第1期82-84,共3页Orthopedic Journal of China

基　　金：湖北省科委重点科技研究项目 (No 2 0 0 2P1 50 3)

摘　　要：目的 :探索从人骨肉瘤细胞条件培养基中提取骨形态蛋白 (BMP)的方法 ,并测定其生物学活性。方法 :收集人骨肉瘤细胞条件培养基 ,通过浓缩、透析 ,SephcrylS -10 0凝胶层析纯化 ,BMP单克隆抗体鉴定所需洗脱峰 ,SDS PAGE测定分子量 ,小鼠肌袋实验检测其骨诱导活性。结果 :BMP单抗鉴定所提蛋白为BMP ,SDS PAGE显示分子量约为 2 1kD ,能够在小鼠肌肉内产生异位骨化。结论 :人骨肉瘤细胞条件培养基中含有BMP ,分离后具有良好的生物学活性 ,而骨肉瘤细胞可以在体外长期培养生长 ,为BMP的大量提取、临床应用提供一个有益的方法。Objective :To find out a method of extraction and purification of bone morphogenetic protein (BMP) from the conditioned medium of human osteosarcoma cell,and evaluate its biological activity.Methods:Conditioned medium of human osteosarcoma cell was collected,concentrated and dialyzed.The concentrated protein was purified through gel filtration chromatography on Sephcryl-S-100.The purified protein was tested by BMP monoclonal antibody (McAb),its molecular weight (MW) was determined by SDS-PAGE and its biological activity was demonstrated by heterotopic ossification.Results:The purified protein was proved to be BMP by BMP McAb,and have a satisfactory heterotopic ossification,its MW was about 21 kD.Conclusion:BMP is contained in the conditioned medium of human osteosarcoma cell and have a satisfactory biological activity after certain purification.Because the osteosarcoma cell can be cultured and grow for a long time in vitro,this method will be helpful to a vast extraction of BMP and clinical application.

关 键 词：骨肉瘤细胞 培养基 骨形态蛋白 蛋白质纯化 细胞培养 

分 类 号：R738.1[医药卫生—肿瘤]