不同纯化方法对大鼠胰岛收获效果的研究.docVIP

不同纯化方法对大鼠胰岛收获效果的研究 作者：吴钢 作者单位：复旦大学附属华山医院 普外科，上海 200040 【摘要】 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（P0.01）。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（P0.01）。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。 【关键词】 胰岛细胞 分离 纯化 乌司他丁 大鼠 Comparative study on the harvest effect of islets of Langerhans with different purified methods WU Gang， CAI Duan， WU Jianhua，et al. Department of General Surgery， Huashan Hospital， Fudan University， Shanghai 200040 Abstract 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 （P0.01）. The yield of islet in the groups that purified with hand-pick under optical microscopy was significantly higher than that groups with Ficoll-400 （P0.01）. Conclusion Purification with hand-pick under optical microscopy can