Iodixanol density gradient preparation in university of wisconsin solution for porcine islet purification.
Generally, prior to the purification of isolated pancreatic islets, the collagenase-digested tissue is incubated in the University of Wisconsin solution (UWS; approximately 320 mOsm) for osmotic stabilization to preserve or improve the density differences between islets and acinar fragments. The adverse effects arising from the subsequent pelleting and resuspension of the islets in a second, different (often highly hyperosmotic) purification solution are avoided in the protocol described here; preparation of the purification medium is simply achieved by mixing the UWS preincubated islets with a second UWS containing the inert impermeant iodixanol. Flotation of the islets isolated from juvenile porcine pancreases through this mildly hypertonic (approximately 380 mOsm) gradient of iodixanol-UWS achieves a much higher recovery of islets of an improved viability than the customary method using a Ficoll gradient. The method has been extended to human islet purification.
van der Burg
Michael P M
MP
Department of Surgery K6-50, Leiden University Medical Centre, Leiden, Netherlands. burg@lumc.nl
Graham
John M
JM
D000818:Animals; D002499:Centrifugation, Density Gradient / Q000379:methods*; D007515:Islets of Langerhans / Q000378:metabolism*; D012996:Solutions; D013552:Swine; D014283:Triiodobenzoic Acids / Q000737:chemistry*; D014495:Universities; D014922:Wisconsin
D012996:Solutions; D014283:Triiodobenzoic Acids; C044834:iodixanol