分离纯化小鼠胰岛的方法学研究

目的：探讨分离纯化小鼠胰岛的方法。方法将10～20周、体质量25～30 g 的雄性 ICR 小鼠断颈处死后，经十二指肠大乳头，进入胰管行胶原酶-P 灌注、原位消化；然后在显微镜下挑取胰岛；采用台盼蓝染色、DTZ 染色，以及葡萄糖、氯化钾刺激胰岛素分泌等方法观察胰岛的活性、纯度及功能。结果每只小鼠分离的胰岛可达200～250个，且纯度达99％、活性大于98％，20 mmol·L－1葡萄糖或40 mmol·L－1氯化钾刺激胰岛后胰岛素的分泌是基础分泌量的4～7倍。结论本方法可分离纯化到活性、纯度及生物活性高的胰岛。

A Method for Isolation and Purification of Mouse Pancreatic Islets

Objective To explore a method for isolation and purification of mouse pancreatic is-lets.Methods Male ICR mice weighing 25-30 g （10-20 weeks of age）were euthanized by cervical dislocation and collagenase P was injected into pancreatic duct through the major duodenal papilla for in suit digestion.The islets were collected by hand-picking under the microscope.The viabili-ty,purity and function of islets were assessed by trypan blue staining,DTZ staining,and insulin secretion stimulated by 20 mmol·L-1 glucose or 40 mmol·L-1 KCl.Results A total of 200-250 pancreatic islets were obtained from each mouse with a purity of 99% and a viability of greater than 98%.The basal insulin secretion was increased 4-7-folds by stimulation with 20 mmol·L-1 glucose or 40 mmol·L-1 KCl.Conclusion The method can be used to obtain mouse pancreatic islets with high viability,purity and bioactivity.