van der Burg Michael P M, Graham John M
Department of Surgery K6-50, Leiden University Medical Centre, Leiden, Netherlands.
ScientificWorldJournal. 2003 Dec 1;3:1154-9. doi: 10.1100/tsw.2003.107.
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.
一般来说，在纯化分离的胰岛之前，将胶原酶消化的组织置于威斯康星大学溶液（UWS；约320 mOsm）中孵育以进行渗透稳定处理，以保持或改善胰岛与腺泡碎片之间的密度差异。在此处描述的方案中，避免了随后将胰岛在第二种不同的（通常是高渗的）纯化溶液中沉淀和重悬所产生的不利影响；纯化培养基的制备只需将预先用UWS孵育的胰岛与含有惰性非渗透性碘克沙醇的第二种UWS混合即可。通过这种轻度高渗（约380 mOsm）的碘克沙醇-UWS梯度对幼年猪胰腺分离的胰岛进行浮选，与使用Ficoll梯度的传统方法相比，可实现更高的胰岛回收率和更好的活力。该方法已扩展到人类胰岛纯化。
