两种酶对猪胰岛消化分离效果的对比

The effective comparison of adult porcine islet isolation by two kinds of enzymatic digestion

目的 研究单纯酶 (胶原酶Ⅳ )与复合酶 (由胶原酶Ⅰ、Ⅳ、弹性蛋白酶、纤维素酶组成 )对成年猪胰岛的分离效果。方法 采用经胰管灌注消化酶的方法分离胰岛 ,用双硫腙染色做胰岛计数 ,台盼蓝染色判定胰岛活化水平 ,胰岛培养第 2d、第 4d、第 6d测定胰岛素含量 ,培养第 6d做胰岛活性评价 ,电镜观察胰岛的形态结构。结果 经复合酶消化法制备的胰岛数量为 (4 915±l0 42 )个胰岛 /克胰腺 ;而经单纯酶消化的胰岛数量为 (30 12± 989)个胰岛 /克胰腺。两组比较差异显著 (P <0 .0 5 )。结论 复合酶消化法是一种优于单纯酶消化法的成年猪胰岛分离方法。

Objective To study the effects of adult porcine islet isolation by a single enzymatic solution(SES) and a multi enzymatic solution(MES). Methods The islets was isolated by the perfusion of two solutions via pancreatic duct. The islets were counted by dithizone staining. The standard of isolated live cells was justified by trypan blue staining. The insulin content in the culture medium and the islet activity were determined on the 2nd day, 4th day, 6th day of the culture. The morphology of the islets was observed under an electron microscope. Results The islets counted by MES ( 4?915 ± 1?042 ) islets/g pancreas were significantly more than those by SES ( 3?012 ±989) islets/g pancreas ( P < 0.05 ). No significant difference was found in islet cell morphology or function between MES and SES ( P > 0.05 ). Conclusion The method of multi enzymatic digestion was superior to that of single enzymatic digestion on adult porcine islet isolation.