Semi-automatic Isolation and Purification of Adult Pig Islet Cells

Objective To establish effective method for large-scale purification of islet cells from pig pancreas.Methods Pig pancreas tissue was digested with collagenase P through continuous infusion.Pig islets was purified by Ficoll discontinuous density gradient centrifugation medium.Dithizone(DTZ)staining was used to observe the morphology of islet cells clusters(ICCS),to count islet equivalent IEQ,and to detect the purity of the islet.AO-PI staining was used to detect islet viability and the islet function assessed by insulin release assay in vitro.Results The number of the islets collected was(4 387±617)IEQ/g in average from pancreas before purification,and(3 192±756)IEQ/g pancreas after purification.The average purity of islets cells was(60.15±2.35)%,and the motilityrate was(80.00 ± 0.47)%.Conclusions The islet cells obtained by this method have better purity and good biological activity.That's to say the Semi-automatic isolation and purification of small-eared pig islet cells is successful and feasible.The established method can be applied in large-scale purification of fully functional islet cells from pig pancreas,and can meet the need for further studies of xenotransplantation.