深低温保存对气管组织细胞活力的影响

Effects of cryopreservation on the viability of tracheal tissue and cell in rats

目的探讨深低温保存对气管组织细胞活力的影响程度。方法切取SD大鼠气管后立即放入含有新鲜配置的低钾右旋糖酐(LPD)溶液的冻存管,在程序降温仪降至-80℃后投入液氮中保存,分别保存24 h、15 d、30 d、60 d、120 d。然后对气管组织体外培养,加入3H-TdR以做标记,最后使用β液体闪烁计数器检测组织细胞吸收情况。结果与冷冻前比较,低温保存后气管组织细胞3H-TdR掺入率降至75.3%～81%。冷冻15 d以后3H-TdR掺入率差异无统计学意义(P>0.05)。结论深低温保存后气管组织细胞保留了70%～80%的活力,损伤主要发生在冷冻早期。采用3H-TdR体外组织培养是检测气管组织细胞活力的有效方法。

Objective To investigate the effects of cryopreservation on the viability of tracheal tissue and cell in rats.Methods The tracheas from SD rats were removed and immersed immediately in low potassium dextran（LPD） solution.Then the sterile plastic tubes were sealed,and frozen up to-80℃ in a programmable freezer.The tubes were stored in liquid nitrogen for different time of preservation（24h,15d,30d,60d,120d）.Then the specimens were cultured and labeled with 3H-TdR in vitro in order to check the absorption of them, and the cells were counted by liquid scintillation counter. Results The average 3H-TdR incorporation in the trachea after cryopreservation was decreased by 75.3% - 81.0%, as compared with that before cryopreservation. There were no significant differences in 3H-TdR incorporation 15d after eryopreservation among the groups. Conclusion After cryopreservation,the tissue and cells of trachea can maintain 70% - 80% of viability. The damage of tracheal viability mostly occurs in the early stage of freezing. The measurement of 3H-TdR incorporation is a reliable method as a market of the viability of tissue and cells of trachea.