不同年龄兔软骨细胞培养和形态学研究  

The Study on Chondrocyte Culture and Morphology of Rabbits in Different Ages

作　　者：李昊[1] 

机构地区：[1]山西医科大学第二临床医学院,山西太原030001

出　　处：《山西职工医学院学报》2008年第1期15-17,共3页Journal of Shanxi Medical College for Continuing Education

摘　　要：目的:研究兔关节软骨细胞的分离、培养方法,观察不同年龄兔软骨细胞的形态学特性。方法:无菌条件下,从不同年龄新西兰白兔的四肢关节髁突面削取软骨片,采用机械-酶消化法分离软骨细胞,经台盼蓝拒染计数,传代培养。应用甲苯胺蓝及Ⅱ型胶原免疫组化对细胞进行鉴定,利用相差显微镜及透射电镜观察细胞形态,激光共聚焦扫描显微镜检测软骨细胞直径。结果:培养2 d^3 d,细胞贴壁、变形,呈多角形;8 d左右,细胞融合成层。甲苯胺蓝及Ⅱ型胶原染色阳性。透射电镜观察显示,随兔年龄改变,关节软骨细胞的超微结构也有所差异。用激光扫描共聚焦显微镜对其直径进行检测,显示三组细胞的直径两两之间都有差别,即成年软骨细胞直径大于幼年软骨细胞直径,小于老年软骨细胞直径(P<0.001)。结论:本研究建立了简单易行的软骨细胞分离、培养方法;初代、第2代细胞生长良好,适合于实验研究;不同年龄兔软骨细胞的形态结构有所改变,为了解骨性关节炎的发展变化提供一定依据。Objective： To study the methods for isolation and culture of rabbit articular chondrocyte and observe the morphological characteristics of rabbit chondrocyte in different ages. Methods：Cartilage was harvested under sterile conditions from synovial joints of New Zealand white rabbits in different ages to isolate chondrocyte by mechanical-enzymatic digestion method and to count and subculture the cells by trypan blue exclusion method . Then the cultured cells were identified with toluidine blue staining and Type II collagen immunohistochemistry reaction, the cell morphology was observed by phase-contrast microscope and transmission electron microscope （TEM） and the diameter of the chondrocyte was observed by laser confocal scanning microscope （LCSM）. Results： Cultured cells showed cell attachment , deformation or multi-angle in 2 - 3 days and the cells formed a cohesive multiplayer on plastic surface about 8 days. Toluidine blue and collagen type II were positive. The TEM observation discovered the ultramicrostructure of articular chondrocyte had some differences as the rabbit＇s age changed. There were significantly alternations between the diameter of each chondrocyte in three groups. That is to say, adult chondrocyte diameter is more than juvenile chondrocyte diameter and less than senile chondrocyte diameter （P 〈 0. 001 ）. Conclusions：The study sets up a series of simple and feasible methods for isolation and culture of chondrocyte. The cells of the initial and the second passages grow well and they are suitable for experimental research. The morphosis of the rabbit chondrocyte in different ages has some changes and it can provide the related theoretical basis for the development and changes of osteoarthritis.

关 键 词：不同年龄 关节 软骨 细胞学 超微结构 

分 类 号：R68[医药卫生—骨科学]