大鼠胰岛的分离和胰岛移植

Comparison of different methods for isolation of rat islets and islet transplantation

目的研究大鼠胰岛分离和移植的方法。方法采用胶原酶消化及不纯化的胰岛分离方法。用双硫腙(DTZ)法判定胰岛纯度,台盼蓝排斥试验评估胰岛存活率;SD大鼠胰岛制备物行同种异体胰岛移植后,动态检测受体血糖、胰岛素及C肽变化,并作移植部位形态学观察,以判定其功能。结果2次消化组胰岛收获量和纯度较单次消化组高,与不加甘油单次消化组相比有显著性差异(P<0.05);胰岛活率2次消化组显著高于单次消化组(P<0.05)。在有效的免疫抑制方案下胰岛移植物能长期存活,有良好的胰岛素和C肽分泌功能,可维持正常的血糖;移植后2个月在汇管区可见较多形态完整的胰岛细胞。结论胶原酶2次消化和不纯化的分离方法,可获得较高质量和数量的胰岛移植物,且重复性好。

Objective To study the method for isolation of rat islets and islet transplantation.Methods Isolation of rat islets were performed with collagenase digestion and non-purification. For identification, islet samples were stained with dithizone(DTZ). The viability of islets was determined with trypan blue test. For evaluating function, the concentrations of blood glucose, insulin and C-peptide were determinated developmentally in recipients and the sites of transplantation were observed morphologically after islet samples of SD rats were transplanted by means of a transhepatic portal embolization in Wistar rats.Results The yield and purity of islets in twice digestion group were higher than those in single digestion group and compared with single digestion group without glycerin, the differences of yield and purity were significant(P<0.05). The survival rate of islets in twice digestion group was significantly higher than that in single digestion group(P<0.05). Islet grafts could keep long-term viability with effective immunosuppressive strategy and maintained normal blood glucose, insulin and C-peptide beyond 60 days. There were many morphologically intact islets in liver of recipients two months after transplantation.Conclusion High quality and more islet grafts could be harvested by twice digestion and non-purification. The method had better repetition.