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目的经微重力培养的新鲜和冻存胰岛联合移植提高1型糖尿病的治疗效果。方法将分离纯化的大鼠胰岛分为(1)体外实验组:实验组1.1:冻存的胰岛经微重力培养;实验组2.1:冻存的胰岛在普通培养基中培养;对照组1:新鲜大鼠胰岛经微重力培养。观察胰岛收获率和体外胰岛素分泌情况。(2)胰岛移植组:即将新鲜和冻存的胰岛经微重力或普通培养7 d后分别移植入受体鼠体内,观察移植效果。结果经微重力培养的各组胰岛收获率、DNA含量和胰岛素含量高于普通组。普通培养组在培养后期胰岛素分泌明显下降,并且胰岛素刺激指数明显低于经微重力培养组(P<0.05)。移植经过微重力培养的500 IEQ新鲜胰岛和1 500 IEQ冻存胰岛在移植后1周内可达100%纠正糖尿病,全部受体维持正常血糖耐受曲线一直到观察结束。结论采用细胞内低温保存液(HTS)结合细胞冻存液(DMSO)对大鼠胰岛进行冻存前后经微重力培养可以明显提高胰岛冻存质量,是目前胰岛冻存的最佳选择。使经微重力培养的新鲜和冻存胰岛联合移植一次治愈糖尿病成为可能,并有效的节约了胰岛资源,提高了移植效果。
Objective To improve the therapeutic effect of type 1 diabetes mellitus by co-transplantation of fresh and frozen pancreatic islets cultured by microgravity. Methods Isolation and purification of rat islets were divided into (1) in vitro experimental group: experimental group 1.1: frozen pancreatic islets were cultured by microgravity; experimental group 2.1: frozen pancreatic islets cultured in normal medium; control group 1: fresh Rat islets were cultured by microgravity. Observe the rate of islet harvest and insulin secretion in vitro. (2) Islet transplantation group: The fresh and frozen islets were transplanted into recipient mice after being gravimetrically or routinely cultured for 7 days, and the effect of transplantation was observed. Results The rate of islet harvesting, DNA content and insulin content in all groups cultured by microgravity were higher than those in normal group. Insulin secretion in the normal culture group decreased significantly in the later period of culture, and the insulin stimulation index was significantly lower than that in the microgravity culture group (P <0.05). Transplantation of 500 IEQ fresh islets and 1 500 IEQ cryopreserved grafts with microgravity transplanted 100% correct diabetes within 1 week after transplantation, with all receptors maintaining normal glycemic curves until the end of observation. Conclusion The cryopreservation of pancreatic islets by cryopreservation fluid (HTS) combined with cryopreservation solution (DMSO) before and after cryopreservation of rat pancreatic islets is the best choice for pancreatic islets cryopreservation. It is possible to combine the fresh and cryopreserved pancreatic islets to cure diabetes mellitus after microgravity culture, which effectively saves islet resources and improves the transplantation effect.