Isolation, Purification and Culture of Rat Islet Cells

Objective To develop a method of isolating and purifying and culturing rat islet cells. Methods The isolation of adult rat pancreatic islets was carried out by injection of Hanks into pancreatic duct and digestion of collagenase Ⅴ. A discontinuous Ficoll 400 solution was applied to the purification of the islets, and the viability and purity of islets were determined by trypan blue and dithizone staining. Islets function was evaluated by the glucose stimulating insulin release test. ResultsThe yield of islets was(540±150) islets/pancreas, with viability 90% and purity90%. The insulin from the islets fed with high concentration glucose was 2 times that with low concentration. ConclusionPancreatic ductal Hanks injection and digestion by collagenase Ⅴ, followed by purification of islets with discontinuous Ficoll 400 solution, can reproducibly yield functional and pure islets.