大鼠甲状旁腺细胞玻璃化冻存效果评价

Evaluation of vitrified cryopreservation of parathyroid cells

目的观察玻璃化方法冻存甲状旁腺细胞的效果,探索甲状旁腺细胞经冻存复苏后移植治疗甲状旁腺功减退症的可行性。方法应用常规方法和玻璃化方法分别冷冻保存甲状旁腺细胞,冻存前后,对甲状旁腺细胞的结构和功能进行检测。随机将甲状旁腺功能减退症模型大鼠分为4组,分别移植培养液、未冻存的甲状旁腺细胞、常规方法冻存复苏后的甲状旁腺细胞和玻璃化方法冻存复苏后的甲状旁腺细胞,用血清钙水平的变化来判断移植物的存活。结果冻存前后,细胞的结构和功能没有明显变化,常规方法和玻璃化方法效果相似。移植后大鼠血清钙浓度的水平,4组分别是(1.35±0.05)mmol/L、(2.18±0.12)mmol/L、(2.22±0.09)mmol/L、(2.11±0.11)mmol/L,各实验组分别与移植前比较,血清钙水平的增高有统计学意义(P<0.05),各实验组在移植物存活时间上的差异没有统计学意义(P>0.05)。结论玻璃化方法冷冻保存甲状旁腺细胞有效,细胞复苏后,可应用于移植。

Objective To study the effects of cryopreservation,vitrification of parathyroid cells in rats.To examine if the cryopreserved parathyroid cells transplantation could be performed in treating hypoparathyroidism when the cells have been resuscifed.Methods Parathyroid cells were cryopreserved with routine and vitrification method respectively.Before and after cryopreservation,the structure and function of parathyroid cells were measured respectively.The recipients were divided into 4 groups randomly.Every rat of each group was respectively injected with culture medium,parathyroid cells,cryopreserved parathyroid cells with routine method and cryopreserved parathyroid cells with vitrification method.The changes in levels of serum calcium were measured continuously to determine the graft＇s survival profile.Results There were no significant differences on the structure and function of parathyroid cells after cryopreservation.And it was found that the results with vitrification method were similar to that with routine method.After transplantation the levels of serum calcium in each group were（1.35 ± 0.05） mmol/L,（2.18 ± 0.12） mmol/L,（2.22 ± 0.09） mmol/L and（2.11 ± 0.11） mmol/L.In experiment groups they increased significantly by contrast with those of pretransplantation respectively（P 〈 0.05）.There were no statistical differences of the survival time of transplantation in experiment groups（P 〉 0.05）.Conclusion The vitrification method is effective in cryopreserving parathyroid cells.The cryopreserved parathyroid cells transplantation has practical significance in the treatment of hypoparathyroidism.