阶段性酶消化法高效分离关节软骨细胞

Effective isolation of articular chondrocytes by using staged enzymatic digestion

目的探讨软骨细胞的简便、高效分离方法。方法应用自行设计和制造的细胞消化器,对10只新西兰白兔关节软骨细胞进行阶段性酶消化分离,以45min为一个消化单位时间,计算细胞收获率和细胞存活率。并与传统分离法进行比较。结果应用改进的细胞消化器通过阶段消化分离法可获得大量具有活性的软骨细胞,细胞收获率4877±726×104个/g,细胞存活率98.7±1.6%,比传统消化分离法有明显提高。结论应用自制细胞消化器进行软骨细胞的阶段性酶消化分离简便宜行,可高效分离软骨细胞。

Objective To explore a simple and effective method for isolating chondrocytes. Methods By using our self-designed and manufactured digestion chamber, Cartilages from 10 rabbits were digested by 0.2% collagenase Ⅱ.The harvest rate and viability were observed. The effects between staged enzymatic digestion and traditional method on the harvest and viability of the chondrocytes were analyzed. Results Plenty of activated chondrocytes can be isolated by staged enzymatic digestion, the number of chondrocytes gained from wet rabit cartilage was 4 877 ± 726 × 10^4/g with a viability of 98.7 ±1.6 %, more effective than traditional method significantly. Conclusion Staged enzymatic digestionis the best method which can obtained a great number of chondrocytes but also can maintain good growth activity, our self-designed digestion chamber can make the isolation of chondrocytes more efficient