成人胰岛细胞的分离

目的　探讨成人胰岛细胞的分离及胰腺冷缺血时间与胰岛细胞活率的关系。方法　采取胰、肾联合切取 ,经腹主动脉插管 ,用自制的灌注器以高渗枸橼酸盐嘌呤溶液 (HC A液 ) 15 0 0～2 0 0 0ml进行原位灌洗。将肾与胰腺、十二肠、脾分离 ,采用胶原酶P消化胰腺 ,分离并纯化胰岛细胞 ,以双硫腙及丫腚橙染色 ,测定所得到的胰岛细胞的纯度及活率。结果　胰腺的冷缺血时间为 2 .5～ 8h ,温缺血时间为 0～ 3min。胰岛收获量为 (2 .38± 0 .6 7)× 10 3 IEQ/g ,纯化后胰岛细胞的活率为 19%～ 83% ,冷保存指数 (或输注指数 )为 9.8～ 6 6 .4。胰岛细胞的活率与胰腺的冷缺血时间呈负相关 ,冷保存指数和胰岛细胞活率呈负相关。结论　以高渗枸橼酸盐嘌呤溶液灌洗、胶原酶P消化胰腺所得到的胰岛细胞活率高 ,但胰腺的冷缺血时间不宜超过 5h。

Investigation of the cold ischemia preservation of human pancreases islets

Objective To establish a new method of cutting human pancreases from cadaver donors for islets transplantation, and to discuss the relationship between activity of islets and the cold ischemia preservation time after hypertonic citrate adenine solution perfused.Methods 1500-2000 ml of hypertonic citrate adenine solution was perfused into aorta. Kidey-pancreases-duodenum-spleen were cut together. Digestion of pancreases was done with collagenase P. The islets were isolated, purified and stained. The purity and activity of the islets were measured.Results There was a negative correlation between activity of islets and the cold ischemia preservation time. But there was a positive correlation between activity of islets and the ischemic index.Conclusion The activity of the islets infused with hypertonic citrate adenine solution and degested with collagenase P is high, but the cold ischemia preservation time of the pancreases preserved in hypertonic citrate adenine solution is better not more than 5-6 h.