兔软骨细胞的分离与培养  

The isolation and culture of rabbit's articular chondrocytes

作　　者：李保林 王海彬[2] 周正新[3] 杨荣建[2] 袁浩[2] 

机构地区：[1]广州市越秀区正骨医院,510045 [2]广州中医药大学第一附属医院,510405 [3]安徽中医学院附属医院,230038

出　　处：《广州医药》2005年第1期2-4,共3页Guangzhou Medical Journal

摘　　要：目的　探讨兔软骨细胞培养方法。方法　采用胶原酶Ⅱ分段消化法从 4周龄新西兰兔的关节软骨中分离出软骨细胞并进行原代和传代培养。每日在倒置显微镜下观察细胞形态及其生长情况。并用甲苯胺蓝异染色进行细胞表型鉴定。结果　软骨细胞形成单层的时间原代培养 1周左右 ,传代培养约 4～ 5天 ,细胞以圆形或上皮样细胞形态为主。甲苯胺蓝染色证实细胞可合成蛋白多糖 ,异染反应主要集中在细胞集落样生长区 ,异染程度以原代培养最为显著。结论　采用本方法培养兔软骨细胞是可行的。Objective To study the method of isolation and culture of rabbit's articular chondrocytes. Methods Articular chondrocytes were obtained from the cartilage of New Zealand rabbit aged 4 weeks adopting method of sequential digestion by 0.2% collagenaseⅡ. Primary culture and subculture of chondrocytes were performed respectively. The morphological changes and growth feature were recorded under the inverted microscope each day. Histochemical staining was used to determine proteoglycans synthesis of chondrocytes. Results The time for chondrocytes to develop a monolayer was about one week in primary culture, while four to five days in subculture. Rounded and epithelioid chondrocytes occupied most of cells. The proteoglycans secreted by chondrocytes were stained to red violet by toluidine blue staining. The metachromasia location was confined to large aggregate of rounded cells and multilayered areas. It was more prominent in primary cultures, compared with subculture. Conclusion Using this method to isolate and culture rabbit's articular chondrocytes is feasible.

关 键 词：兔 软骨细胞 分离 培养 细胞培养 胶原酶Ⅱ分段消化法 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]