目的　建立分泌抗人成骨肉瘤单克隆抗体 (mAb)的杂交瘤细胞系 ,并对其分泌的mAb进行鉴定。方法　用成骨肉瘤细胞系 (OSPS 96 0 7)免疫Balb/c小鼠 ,取脾细胞按常规方法融合、筛选、克隆化及腹水mAb制备。取骨肉瘤手术标本 (6 0例 )及正常组织 (6种 ) ,用mAb进行免疫组化ABC染色。结果　筛选出 2株能持续、稳定分泌特异性抗骨肉瘤mAb的杂交瘤细胞株 (6E5和 3F7)。 2株杂交瘤细胞经过 10 0d连续培养 ,分泌mAb的特性稳定、mAb特异性强、效价高。免疫组化染色检测表明 ,mAb 6E5和 3F7针对的抗原 ,在成骨肉瘤组织及成骨肉瘤细胞系均呈高表达 (70 % )。针对mAb 6E5的抗原主要分布于细胞核上 ,针对mAb 3F7的抗原主要分布于细胞浆内。 6种正常组织中除胃粘膜可见弱阳性反应外 ,其余未见明确的阳性反应。结论此两株杂交瘤细胞分泌的mAb对成骨肉瘤细胞具有特异亲和性 ,为成骨肉瘤的早期诊断、免疫治疗及发现成骨肉瘤新的标志物奠定了基础 Objective To establish a hybridoma cell line secreting anti-human osteosarcoma monoclonal antibody (mAb) and identify the secreted mAb. Methods Balb / c mice were immunized with osteogenic sarcoma cell line (OSPS 96 0 7). The spleen cells were fused, selected, cloned and prepared ascites by conventional methods. Osteosarcoma specimens (60 cases) and normal tissues (6 pieces) were obtained. Immunohistochemical ABC staining was performed with mAb. Results Two hybridoma cell lines (6E5 and 3F7) that could consistently and stably secrete specific anti-osteosarcoma mAb were screened out. Two hybridoma cells were cultured continuously for 10 days, the characteristics of secreting mAb were stable, mAb-specific and high titer. Immunohistochemical staining showed that the antigens against mAb 6E5 and 3F7 were overexpressed in both osteogenic sarcoma and osteogenic sarcoma cell lines (70%). Antibodies directed against mAb 6E5 were mainly found in the nucleus, and antigens against mAb 3F7 were mainly located in the cytoplasm. 6 kinds of normal tissue except gastric mucosa can be seen weak positive reaction, the rest did not see a clear positive reaction. Conclusions The mAb secreted by the two hybridoma cells has specific affinity to osteosarcoma cells and lays the foundation for the early diagnosis, immunotherapy and discovery of new osteosarcoma markers for osteosarcoma