两种不同纯化大鼠胰岛细胞方法的比较

Comparison between Two Methods for Purification of Rat Islets

目的比较分析OptiPrep和Ficoll两种介质纯化大鼠胰岛细胞的效果。方法将24只SD大鼠随机分成2组,采用明尼苏达大学改良方法提取大鼠胰岛细胞,消化分离胰岛细胞后,分别采用不连续密度梯度Ficoll离心法和OptiPrep离心法纯化胰岛细胞,比较纯化后胰岛细胞数量、纯度、存活率及功能,观察OptiPrep和Ficoll两种介质对大鼠胰岛细胞纯化效果的影响。结果与Ficoll组相比,OptiPrep组胰岛细胞的产量(分别为304±13.57和346±9.09)和纯度(分别为88.25%±2.22%和94.00%±0.82%)均显著增加(P<0.05);OptiPrep组与Ficoll组胰岛细胞的存活率(分别为93.50%±1.29%和91.50%±1.29%)和功能(胰岛素释放指数分别为3.04±0.05和3.02±0.06)的差异无统计学意义(P>0.05)。结论 OptiPrep法纯化大鼠胰岛细胞比传统的Ficoll法更有优势,而且OptiPrep价格相对便宜,可以作为纯化大鼠胰岛的常规方法。

Objective To compare the effects of OptiPrep and Ficoll on the purification of rat islets.Methods Islets were isolated from the pancreata of 24 SD rats by means of modified University of Minnesota′s method.After an successful enzymatic digestion,the digested tissue was randomly purified by Ficoll and OptiPrep.Compared the islet yield,purity,survival and function between the two groups after purification and the effects of two media on the purification of rat islets.Results Compared with the Ficoll group,yield and purity of islets were significantly higher in the OptiPrep group（304±13.57 vs 346±9.09;88.25% ± 2.22% vs 94.00% ± 0.82%,P 0.05）;there were no significant differences between the two groups in survival and function of islets（93.50% ± 1.29% vs 91.50% ± 1.29%;SI 3.04 ± 0.05 vs 3.02 ± 0.06,P 0.05）.Conclusion The purification method using OptiPrep has some advantages over the traditional purification method using Ficoll.Owing to its cheap price,OptiPrep can be used as an conventional method for purification of rat islets.