浙江医学Issue(24)：2152-2154,3.
小鼠胰岛密度梯度离心方法的研究
Method of gradient centrifugation for mouse pancreatic islet isolation
摘要
Abstract
Objective To establish a stable method for mouse pancreatic islet isolation. Methods Male Balb/c mice were used for islet harvest and isolation. The pancreas were digested by 0.5mg/ml col agenase, and the islets were isolated and puri-fied with different gradient centrifugation strategy, stained with DTZ, and observed by microscopy to determine an optimal proto-col for islet isolation. AO/PI staining was used to detect the activity of isolated islets. Different protocols were compared usingx2 and one- way analysis of variance (ANOVA). Results The optimal conditions used for isolation of pancreatic islets from Balb/c mice were as fol ows:using Ficol - 1.108 (4ml), 1.096 (2ml), 1.069 (2ml), HBSS (2ml), acceleration 1- deceleration 1, 400rpm for 4min transfer to 1800rpm for another 8min. The preliminary yield was (125±39)IEQ, purity was (79.6±10.5)%, then underwent another gradient centrifugation with the same strategy, the purity significantly increased to (96.2±1.4)%, while the yield was stil satisfying. AO/PI staining confirmed good activity of the isolated islets. Conclusion The modified islet isolation method is a better choice to obtain higher purity and considerable numbers of islets.关键词
胰岛/分离纯化/密度梯度离心Key words
Islet/Isolation and purification/Gradient centrifugation引用本文复制引用
张金花,邵益,厉智威,方程,田阳,严盛,郑树森..小鼠胰岛密度梯度离心方法的研究[J].浙江医学,2013,(24):2152-2154,3.基金项目
浙江省科技厅创新团队项目 （）
