目的 介绍一种快速分离纯化小鼠胰岛的方法及进行纯化后胰岛的活性、完整性和胰岛内结构的质量分析.方法 雄性ICR小鼠,采用2 mg/mL胶原酶V灌注和消化,并用Hanks液快速洗涤,用Histopaque(R)-1077和Histopaque(R)-1119密度梯度离心对胰岛进行纯化,用手工方法进行胰岛挑选,DTZ、FD-PI染色鉴定胰岛纯度及其活性,透射电镜观察胰岛内的结构.结果 胰岛开始消化至手工挑选前过程耗时< 25 min,每只小鼠得到胰岛数量为:128±36,当量为:145±42,纯度>90%.透射电镜显示胰岛内部血管仍有损伤.结论 采用此方法可快速得到数量较多、结构较完整的小鼠胰岛,且活性高,为进一步进行胰岛的体外质量研究及体内移植奠定了基础.
Objective To introduce a method about how to isolate and purify islets quickly in mice,and to evaluate the activity,integrity,and the inner structure of the purified islets.Methods ICR mice,male,were chosen in the experiment,and were perfused by collagenase V with the concentration of 2 mg/mL.Then the pancreas was digested and washed by the Hanks solution for three times.The tissues were purified by Histopaque(R)-1077 and Histopaque(R)-1119.And the islets were selected by hand.The DTZ,FD-PI staining was done to assay the purity and activity of the islets.Electron microscope inspection was done to evaluate the inner structure of the islet.Results It takes less than 25 min from islet isolation to choose islets by hand.We got 128 ± 36 islets or 145 ± 42 IE from each mouse,and the purity was >90%.But there were still injuries in the inner structure of islet through TEM.Conclusion Intact and high quality activity islets can be got from the mice by this method,which made a good foundation for the further quality research of islet in vitro and the islet transplantation in vivo.