一步法联合冻存提高大鼠胰岛冻存质量的研究

Improving the quality of cryopreserved islets with co-cryopreserved one step method

目的探讨提高大鼠胰岛冻存质量的方法,以期为建立胰岛库奠定基础。方法将受体鼠分为实验组A组:即一步法联合冻存的胰岛移植组;对照组B组:新鲜的胰岛移植组;对照组C组:逐步法冻存的胰岛移植组。观察3组之间胰岛收获率,活性及移植后效果等方面的差异。结果纯化后收获大鼠胰岛每只(826.87±93.24)IEQ。A组平均胰岛收获率为(87±4)%,高于C组的胰岛收获率(81±4)%。A组和C组经过胰岛刺激素释放实验后均有较B组高的基础胰岛素释放量,但刺激指数则明显低于B组的637.3±39.5。胰岛移植入糖尿病鼠受体后,A组和B组在移植后1周即恢复血糖为正常值,并维持到观察结束。而C组中则需要较长时间纠正血糖到正常。移植(2011.14±114.22)IEQ胰岛在A组和B组可达到100%纠正糖尿病。结论采用全新的细胞内低温保存液(HTS)结合细胞冻存液(DMSO)对大鼠胰岛进行一步法冻存取得了明显优于用DMSO逐步冻存的效果。

Objective To investigate the method to improve the quality of rat islets after cryopreservation to make a basis for establishment of the islet bank. Methods All the recipients were divided into three groups：experiment group A （co-cryopreserved one step islets transplantation） ;control group B （fresh islets transplantation） ;control group C （stepwise cryopreserved islets transplantation） The islet recovery, activity and effect after transplantation among three groups were observed. Results After purification, （826.87 ± 93.24） IEQ per rAt were collected. The mean islet recovery in group A was （87±4） %, higher than （81 ± 4） % in the group C. The basal insulin secretory capacity in group A and group C was higher than that in group B, but the stimulation index was lower than in group B （637.3 ± 39.5）. One week after transplantation, euglycemia was achieved in the group A and group B, and lasted to all the observation time, while in group C, it took longer time to reach euglycemia. Diabetes Mellitus could be cured by （2 011.14 ± 114.22） IEQ islet transplantion in group A and group B. Conclusion The effect using one step method with HTS and DMSO co-cryopreservation was much better than the stepwise method with DMSO.