离心管培养软骨作为半月板移植替代物的可能性研究  

POSSIBILITY OF USING CARTILAGE CULTURED IN CENTRIFUGE TUBE AS A SUBSTITUTE FOR MENISCUS

作　　者：王建[1] 杨志明[1] 解慧琪[1] 秦廷武[1] 

机构地区：[1]四川大学华西医院人类疾病生物治疗教育部重点实验室:干细胞与组织工程研究室

出　　处：《中国修复重建外科杂志》2004年第5期426-430,共5页Chinese Journal of Reparative and Reconstructive Surgery

基　　金：国家重点基础研究发展计划资助项目(973):单一细胞组织工程研究(G1 9990 5430 8)~~

摘　　要：目的　比较培养关节软骨和半月板软骨细胞的生物学特性 ,探讨培养软骨作为半月板移植替代物的可能性。　方法　分别自 3周龄大耳白兔关节软骨和半月板分离软骨细胞 ,行单层传代培养和离心管培养。将离心管培养形成的软骨和 6周龄兔半月板行组织学和透射电镜观察 ,比较关节软骨细胞和半月板纤维软骨细胞的生长曲线。流式细胞术检测第 2、4代关节软骨细胞和半月板纤维软骨细胞周期。　结果　第 4代关节软骨细胞呈去分化 ,似成纤维细胞。离心管关节软骨细胞培养能形成软骨 ,半月板纤维软骨细胞不能形成软骨。培养软骨和半月板的组织学及超微结构差异显著 ,培养软骨中软骨细胞 5 %呈现凋亡。第 2、4代关节软骨细胞中亚二倍体细胞比例明显多于第 2、4代半月板纤维软骨细胞 ( P<0 .0 5 )。　结论　半月板来源的软骨细胞经离心管培养不能形成软骨组织 ,关节软骨细胞离心培养形成软骨不能作为半月板的移植物 。Objective To compare biological characteristics between articular chondrocyte and meniscal fibrochondrocyte cultured in vitro and to investigate the possibility of using cultured cartilage as a substitute for meniscus.Methods Chondrocytes isolated from articular cartilage and meniscus of rabbits aged 3 weeks were respectively passaged in monolayer and cultured in centrifuge tube. Cartilages cultured in centrifuge tube and meniscus of rabbit aged 6 weeks were detected by histological examination and transmission electron microscopy. Growth curves of articular chondrocytes and meniscal fibrochondrocytes were compared; meanwhile, cell cycles of articular chondrocytes and meniscal fibrochondrocytes in passage 2 and 4 were separately measured by flow cytometry.Results Articular chondrocytes in passage 4 were dedifferentiated. Articular chondrocytes formed cartilage 2 weeks after cultivation in centrifuge tube, but meniscal fibrochondrocytes could not generate cartilage. The differences in ultrastructure and histology obviously existed between cultured cartilage and meniscus; moreover, apoptosis of chondrocytes appeared in cultured cartilage. Proportion of subdiploid cells in articular chondrocytes passage 2 and 4 was markedly higher than that in passage 2 and 4 fibrochondrocytes( P <0.05).Conclusion Meniscal fibrochondrocytes can not form cartilage after cultivation in centrifuge tube, while cartilage cultured in centrifuge tube from articular chondrocytes can not be used as graft material for meniscus. Articular cartilage is markedly different from meniscus.

关 键 词：半月板移植 关节软骨细胞 离心管 培养软骨 软骨组织 组织学 养形 传代培养 去分化 生长曲线 

分 类 号：R318.0[医药卫生—生物医学工程]