离心管内培养第1代骺板软骨细胞作为软骨移植材料的可行性

目的：作者前期研究已证明，第1代骺软骨细胞可在离心管内形成富含软骨特异性蛋白基质的软骨组织。本实验进一步观察第1代骺板软骨细胞在离心管内培养形成的软骨组织特征，并与半月板、骺板及关节软骨组织进行比较。方法：实验于2006-07/2007-04在四川大学华西医院科研楼组织工程实验室完成。取4周龄新西兰大白兔四肢骺板分离软骨细胞，行单层传代培养，取第1代细胞行无支架离心管培养，观察其形成的软骨组织特征,另取6周龄新西兰大白兔股骨髁关节软骨、胫骨近端骺板及半月板，与离心管培养软骨一并行透射电镜观察，比较其软骨细胞及细胞外基质结构。结果：第1代骺板软骨细胞经无支架离心管培养能形成软骨。离心管内培养形成的软骨具有独特的超微结构，与骺板及关节软骨有一定的相似性，而与半月板有明显区别。4种细胞都有各自不同的细胞及细胞外基质特征。结论：第1代骺板软骨细胞在离心管内培养形成的软骨组织具有与骺板及关节软骨相似的微观结构。

Feasibility of first passage chondrocytes from epiphyseal plate cultured in centrifuge tube as a substitute for cartilaginous tissue transplantation

AIM: Previous studies of the authors confirmed that the first passage chondrocytes from epiphyseal plate could form cartilaginous tissue containing cartilage specific protein matrix. The present study aimed to investigate the feature of cartilaginous tissue formed by first passage chondrocytes from epiphyseal plate cultured in centrifuge tube, and compare to meniscus, epiphyseal plate and arthrodial cartilage. METHODS: Experiments were performed at the Laboratory of Tissue Engineering of Scientific Research Building of West China Hospital of Sichuan University from July 2006 to April 2007. Chondrocytes isolated from epiphyseal plate of New Zealand rabbits aged 4 weeks were passaged in monolayer and first passage chondrocytes were cultured in centrifuge tube and were observed the possibility of formation of cartilaginous tissue. Articular cartilage of femur condyles, epiphyseal plate of proximal tibia and meniscus were collected from a 6-week-old New Zealand rabbit. The ultrastructure of chondrocytes and extracellular matrix in the three kinds of cartilage and the cultured cartilaginous tissue were observed by transmission electron microscope. RESULTS: The first passage chondrocytes from epiphyseal plate formed cartilaginous tissue in centrifuge tube. Cartilaginous tissue cultured in centrifuge tube possessed unique ultrastructure and was similar to articular cartilage and epiphyseal plate, but it was markedly different from meniscus. Four kinds of cells had different cell and extracellular matrix characteristics. CONCLUSION: The first passage chondrocytes from epiphyseal plate formed cartilaginous tissue after cultivation in centrifuge tube has similar microstructure as epiphyseal plate chondrocyte.