摘要
目的建立一种高效、稳定的大鼠胰岛分离纯化方法。方法选取41只体质量150—380g的成年雄性sD大鼠，按体质量分成4组，采用改进后的方法进行胰岛分离纯化，双硫腙染色计数胰岛、鉴定纯度，吖啶橙-碘化丙锭染色判断胰岛活性，葡萄糖刺激实验检测胰岛功能。结果41只大鼠的胰岛产率为（1056±357）爪／只，各组间产率差异无统计学意义[A组（932±542）个／只，B组（1036±339）个／只，c组（1142±305）个／只，D组（1108±178）爪／只，P〉0．05]。胰岛纯度在90％以上，活率超过90％，胰岛刺激指数为2．08±0．10。结论应用改进的分离纯化方法，能在不同体质量水平大鼠间获得稳定的胰岛高产率，胰岛的纯度、活性和功能可靠。
Objective To set up an effective and stabile method for isolation and purification of rat islets. Methods Forty-one adult male SD rats weighing 150-380 g were used as organ donors. All donors were divided into 4 groups by body weight. Islets were isolated using digestion at 37℃ by ductal injection of collagenase with sampling timely to determine when to stop digestion, and purified by centrifugation of discontinuous Ficoll density gradient. The identification of purified islets was evaluated by dithizone stai- ning. The viability of islet was assessed by fluorescence staining of aridine orange and propidium iodide. The function of purified islets was determined by glucose-stimulated insulin release test. Results The total number of purified islets in one pancreas was 1056 ~ 357. There was no significant difference among the is- let yields of the 4 groups with different body weight （ for groups A, B, C and D ： 932±542, 1036±339, 1142±305, and 1108±178, respectively,P 〉0. 05 ）. The purity of islets was more than 90%. The viability of islets was more than 90%. The simulation index was 2.08±0. 10. Conclusion Using the modi-fied method, the islet isolated and purified from rats with different weight had stable high yield, high puri- ty, high viability and good function.
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2013年第6期1262-1263,共2页
Chinese Journal of Experimental Surgery
关键词
胰岛
分离
纯化
大鼠
Islet
Isolation
Purification
Rat
