关节软骨细胞分离——胶原酶的细胞毒性作用初探

Enzymatic isolation of chondrocytes:the study of cytotoxicity of Collagenase.

目的 :探讨胶原酶的细胞毒性作用 ,以期获得关节软骨细胞的最佳分离效果。方法 :以关节软骨细胞酶消化过程中的细胞收获量、细胞存活率和细胞形态为观察指标 ,采用等密度梯度离心方法 ,比较和检测不同批号的胶原酶随消化时间的延长对软骨细胞活性的影响。结果 :胶原酶对于软骨细胞具有细胞毒性作用 ,且此作用随消化时间的延长而累加。结论 :软骨细胞的酶消化分离过程应采用阶段性消化分离法 。

Viability and integrity of cells are two essential prerequisites in the study of cellular biology after enzymatic digestion.Articular cartilage disaggregation by collagenase digestion has been used successfully to isolate chondrocytes. However, this collagenase damages the cells during the digestion of the cartilagenous, tissue.We have used cell density,as measured by isopycnic centrifugation on polysucrose gradients, as an indicator of cell damage. During the process of isolation,cell viability and morphology were observed in various periods of digestion using different batch collagenases.To obviate the enzymatic cytotoxicity,the method of digestion by stages has been proposed in order to get the best effect of isolation.