不同纯化方法对大鼠胰岛收获效果的研究

Comparative study on the harvest effect of islets of Langerhans with different purified methods

目的探讨不同纯化方法对提高大鼠胰岛收获效果的作用。方法随机选取SD雄性大鼠20只,按胰腺消化和胰岛纯化的方法的不同分为4组,消化过程中胆总管灌注胶原酶V,每组5只:①A组:Ficoll-400法纯化胰岛。②B组:光镜下移液吸移管手工挑拣纯化(手工法)胰岛。③C组:胰腺消化过程加入乌司他丁5000 U/L,Ficoll-400法纯化胰岛。④D组:胰腺消化过程加入乌司他丁5000 U/L,手工法纯化胰岛。结果纯化后胰岛的收获量分别为A组(122±25.30)个,B组(179±21.02)个,C组(187±24.89)个和D组(276±24.85)个。乌司他丁灌注组(C组和D组)胰岛细胞的收获量与相应纯化方法的无乌司他丁灌注组(A组和B组)差异有统计学意义(P<0.01)。手工法纯化组(B组和D组)胰岛的收获量与Ficoll-400纯化组(A组和C组)差异有统计学意义(P<0.01)。结论手工法可明显提高胰岛的收获量,乌司他丁灌注胰腺对大鼠胰岛分离具有保护作用。

Objective To explore how to improve the yield and quality of rat islets with different purified methods. Methods Twenty male SD rats were randomly divided into 4 groups based on different methods of purification. Islets were isolated with pancreatic ductal injection of collagenase V. Group A： Islets were purified with Ficoll-400. Group B： Islets were purified with hand-pick under optical microscopy. Group C： Islets were isolated with pancreatic ductal injection of collagenase V and Ulinastatin and purified with Ficoll--400. Group D： Islets were isolated with pancreatic ductal injection of collagenase V and Ulinastatin and purified with hand-pick under optical microscopy. Results The yield of islet in these four groups was 122±25.30, 179±21.02, 187±24.89 and 276±24.85, respectively. The yield of islet in the groups that isolated with pancreatic ductal injection of collagenase V and Ulinastatin was significantly higher than those groups without Ulinastatin （P〈0.01）. The yield of islet in the groups that purified with hand-pick under optical microscopy was significantly higher than that groups with Ficoll-400 （P〈0.01）. Conclusion Purification with hand-pick under optical microscopy can obtain more islets. Ulinastatin perfusion can protect the islet from trypsin digestion in the process of separation.