冷冻保存对软骨细胞存活率及代谢活性的影响

Effect of cryopreservation on articular cartilage

目的了解各种冷冻保存方法对软骨细胞的存活率和代谢活性的影响,寻找满意的软骨组织冻存方法。方法采用梯度慢速降温法和连续慢速降温法对兔关节软骨进行低温冷冻保存处理,通过荧光染色及35SO4摄入率了解冻存后软骨细胞的存活率和代谢活性。结果采用梯度降温法的软骨细胞存活率为61%,显著高于连续降温法;冷冻保存对软骨细胞的代谢活性有一定的影响,但与对照组的差异并无统计学意义。结论梯度降温法较传统保存方法能显著提高冷冻保存后软骨细胞的存活率,并能维持软骨细胞的代谢活性,是理想的关节软骨冷冻保存方法。

Objective To study the effects of different cryopreservation on the viability and metabolism of chondrocytes so as to find out the ideal method for cryopreservation of articular cartilage.Methods Full-thickness articular cartilage,after pretreated with cryoprotective agents,were cryopreserved using step-cooling or constant cooling protocol.Chondrocyte viability and metabolic activity were assessed using paravital staining and radioactive ~ 35 S-sulfate uptake after thawing.Results Chondrocyte viability differed significantly between fresh cartilage and those that had been cryopreserved.The step-cooling protocol resulted in a better recovery of chondrocytes (around 61%) than constant-cooling protocol.But the metabolic activity per viable chondrocyte in two groups was compared with fresh cartilage.Conclusion Using a step-cooling cryopreservation protocol can get a substantial improvement in chondrocyte survival and don't significantly affect the metabolic activity of chondrocyte.It is an optimal cryopreservation protocol of articular cartilage.