正常人髋关节滑膜细胞有限细胞系的建立及其生物学特性的研究  

Establishment of finite synoviocyte line of normal human hip joint

作　　者：蔡諝[1] 王继芳[1] 卢世璧[1] 黄靖香[2] 顾铮[2] 张曙光[2] 侯宁[2] 

机构地区：[1]解放军总医院骨科,北京100853 [2]解放军总医院病理科,北京100853

出　　处：《中华骨科杂志》2000年第7期435-439,共5页Chinese Journal of Orthopaedics

基　　金：国家自然科学基金资助项目!(39800161)

摘　　要：目的 建立正常人髋关节滑膜细胞系，为进一步研究磨损颗粒在骨－假体界面的作用机制提供体外实验模型。方法 从正常人髋关节滑膜取材，纱巾组织块法原代培养， 1∶ 2传代培养，进行活细胞形态、组织学和电镜观察，测定细胞增殖及活性参数，鉴定细胞染色体，以鼠抗人 CD68单克隆抗体做免疫组化染色 (SABC)，分析细胞类型。结果 原代培养 14 d滑膜细胞从组织块边缘逸出， 21 d密集生长开始传代。传代细胞 12 h内完全贴壁，分裂指数第 4 d最高，然后回落，细胞数量逐渐增加，第 7 d达高峰，倍增时间为 25.3 h。传至 21代，细胞生长及特性稳定，染色体正常。第 5代冻存细胞复苏良好。细胞系由形态、核型和超微结构不同的大、小两种细胞构成，其中小型细胞 CD 68阳性，占 23.17％± 5.60％ (x± s)。结论 体外培养可获得生物学特性稳定的正常人髋关节滑膜细胞系，并保留巨噬细胞样细胞 (MCs)和成纤维细胞样细胞 (FCs)，其不同的形态特征为镜下区分提供了依据，是研究磨损颗粒作用机制较好的体外模型。Objective To establish a finite synoviocyte system of normal human hip joint as an in vitro model for the study of wear particles. Methods Synovial membrane was obtained from the normal hip joint of a 64－ year－ old man and was primarily cultured by gauze kerchief technique. After primary synoviocytes were obtained, the cells were subcultured to the secondary generation. Living cell configuration, histologic and ultrastructural feature of synoviocyte was observed. The proliferation and active parameter were also determined. CD68 monoclonal antibody was used to distinguish different kinds of cells in the system (SABC technique). Results 14 days after primary culture, synoviocyte began to move out from the edge of synovium tissue. The cells began to grow thickly after 21 days and then subculture was performed. The subcultured cells completely anchored in 12 hours. Mitotic index was highest at the 4 days and then declined. The amount of cells increased gradually and arrived at the peak at the 7th day. The doubling time was 25.3 hours. The feature of cells remained stable till 21st passage and the chromosome analysis was normal. The cell line consisted of two kinds of cells with different configurations; the ultrastructure showed the small one being CD68 positive occupying 23.17％± 5.60％ (x± s). Conclusion Finite synoviocyte line of normal human hip joint can be established in vitro with stable biological feature. The macrophage like cells (MCs) and fibroblast like cells (FCs) can be reserved in the system and can be distinguished by different configuration feature. The synoviocyte system is a preferable in vitro model in the study of wear particles.

关 键 词：生物学 人工关节 髋关节滑膜细胞系 体外培养 

分 类 号：R684[医药卫生—骨科学]