摘要
目的探讨大鼠关节软骨细胞培养方法。方法采用胶原酶NB4消化法从1周龄SD大鼠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HE、甲苯胺蓝染色和Ⅱ型胶原免疫组织化学染色进行鉴定。结果软骨细胞形成单层的时间在原代培养1周左右,5d左右即可传代。第5代后部分变为梭形,第7代后绝大部分变为长梭形,增殖能力减弱。结论采用此法可在短时间内获得大量纯化的大鼠软骨细胞。
Objective To study the method of isolation and culture of articular chondrocytes from rat. Methods Articular chondrocytes were obtained from the cartilage of SD rat aged 1 week digested by collagenase NB4. Primary culture and subculture of chondrocytes were conducted. The morphological changes and growth feature were recorded under the inverted microscope each day. The chondrocytes were biologically confirmed by HE staining, toluidine blue and immunocytochemical staining of type Ⅱ collagenase. Results The time for chondrocytes to develop a monolayer was one week in primary culture,while five days in subculture. Some of the cells of the 5th generation turned to spindle-shape,after the 7th generation,the cells became long spindle-shape. The ability of proliferation after 7th generation was weaker. Conclusion The method to isolate and culture of articular chondrocytes in SD rat can acquire pure and enough cartilage cells in short time.
出处
《河北医科大学学报》
CAS
2008年第4期534-536,I0010,共4页
Journal of Hebei Medical University
关键词
软骨细胞
细胞培养技术
大鼠
chondrocytes
cell culture techniques
rats
