兔关节软骨细胞的获取及培养

Obtainment and culture of rabbit articular cartilage

试验采用组织块法和酶消化法获取兔关节软骨细胞,并对这两种方法取得的效果进行对比分析。在酶消化方法上,运用Ham-F12培养液和无Ca2+、Mg2+的PBS液配制2g/L的Ⅱ型胶原酶进行消化,同时结合酶阶段性消化法收获软骨细胞,对细胞成活率进行测定。对最终收获的细胞进行培养观察,测定软骨细胞的分泌活性。结果表明:在获取细胞的数量上,酶消化法明显优于组织块法;在成活率比较试验中,Ham-F12配制组明显高于常规的PBS液(无Ca2+、Mg2+)配制组,两组之间存在显著差异(P<0.05);对体外培养的软骨细胞进行鉴定,结果显示软骨细胞仍具有较强的分泌基质能力。在此试验消化分离体系下,可得到大量的、高活性的软骨细胞,且在体外能维持良好的生长特性。

The aim of this study was to seek an ideal isolation method of rabbit articular chondrocytes. Cells were collected by the means of explant culture and enzyme digestion. Collagenase type Ⅱ confected by serum - free Ham - F12 medium and PBS without Ca^2＋ , Mg^2＋ respectively were used to digest and isolate chondrocytes. Cells were obtained by stages in the process of digestion, which viability were assessed and final cutured for short term in vitro. Result shew： the number of chondrocyte of enzyme digestion was more than that of explant culture ,The viability of chondrocytes isolated with collagenase type Ⅱ confected by serum-free medium were markedly higher than that of PBS （ free of Ca^2＋ , Mg^2 ＋ ）. There was statistical discrepancy between the two enzyme digestion groups（ P 〈0.05 ）. Chondrocytes were polygon and bad capability secreting of collagen type II and GAGs in vitro. In conclusion , large numbers of cbondrocytes were obtained and could keep cultivated properties in vitro with collagenase type Ⅱ confected by serum - free Ham - FI2 medium.