Isolation and purification of pig islet cells

VernacularTitle:猪胰岛的分离与纯化
Author: Anyi QIAO ; Wenhong ZHANG ; Xinjie CHEN ; Shounan YI ; Yide OIAN ; Guohong XIAO ; Xuehu XU ; Yangxi HU
Publication Type:Journal Article
Keywords: pancreatic islets; isolation; purification; islet transplantation; diabetes mellitns
From: International Journal of Surgery 2009;36(5):297-299,封3
CountryChina
Language:Chinese
Abstract: Objective To establish effective method for large-scale purification of islet cells from pig pan-cress. Methods Pig pancreas tissue was digested with collagenase P followed by purification in a HCA-Fi-coil dis continuous gradient using Cobe2991 cell separator. After isolation, the islet cell yield and purity were evaluated with light microscope with DTZ staining, and the islet function assessed by insulin release as-say in vitro. Results The number of the islets coll ected from each pancreas averaged (275 000±20 895)islet equivalents (IEQ) before purification, and (230 350±26 679) IEQ after the purification with discon-tinuous gradient centrifugation. From each gram of the pancreatic tissue, (2710±229) IEQ were obtained with an average purity of (50.2±1.95) %. The purified islets responded well to high-concentration (16.7 mmol/L) glucose stimulation with a 4. 74-fold increase of insulin secretion over the basal level (3.3 mmol/L, P <0.001). Conclusion The established method can be applicable for large-scale purifi-cation of fully functional islet cells from pig pancreas.