中文摘要：

目的：探讨并改良肠道黏膜固有层淋巴细胞（LPL）分离方法。方法：参照Sanos等方法进行改进。用分离液和消化液处理肠组织片段后,通过100μm的尼龙滤网过滤,行零加减速密度梯度离心,观察分析细胞获得率、细胞活率和细胞纯度。结果：每只小鼠肠道LPL的获得量为（5.5±1.7）×106个,细胞活力〉92%;细胞平均直径为（7.5±0.8）μm,细胞平均圆度为0.93±0.03,细胞结团率为（0.4±0.2）%;流式细胞仪检测细胞活率为（94.5±3.2）%,CD4＋T细胞的含量为（72.5±5.2）%。结论：与国内外其他分离方法相比较,该方法简化、高效、稳定,且获取的LPL的细胞数量多、活率高、纯度高,可为肠道黏膜免疫的研究提供可靠的方法,值得推广。

英文摘要：

Objective：To explore and modify the isolation method for mouse intestinal lamina propria lymphocytes.Methods：The isolation method was based on the methods of Stephanie L.Sanos with some improvements.After incubated with separation solution and digestive solution,the intestinal pieces filtered through a gray-mesh（100μm）,and treated with density gradient centrifuging without acceleration and brake.The acquired cell rate,viability and purity of intestinal lamina propria lymphocytes were observed and analyzed.Results：The number of acquired cells was（5.5±1.7）×106 per mouse,and the viability of acquired cells was more than 92%.The average diameter of acquired cells was（7.5±0.8）μm,the average compactness of acquired cells was0.93± 0.03,and the aggregate rate of acquired cells was（0.4±0.2）%.Flow cytometry showed that the viability of acquired cells was（94.50±3.20）%,and the purity of acquired cells stained with CD4was（72.5±5.2）%.Conclusion：Compared with other reported isolation methods,the modified method is simple,efficient and stable,and could acquire sufficient cells with high viability and high purity.Therefore,the modified method provides a reliable method for in-testinal mucosal immune system studies.