Simple method for the collection of large numbers of viable pancreatic islets from normal rats is described. This method involves collagenase digestion of the pancreas, followed by the use of Ficoll-Conray discontinuous gradient for the separation of isolated islets from unwanted acinal debris.
Ficoll-Conray A solution was prepared by mixing undialyzed 12.5% Ficoll and 33.4% Conray in the ratio 2: 1, to make a specific gravity of 1.095 and osmolarity of 396 mOsm/l. The solution B, C, and D, with specific gravities 1.084, 1.072, and 1.048, respectively were obtained by diluting A solution with distilled water.
Using Ficoll-Conray gradient, about200viable islets which maintained excellently their morphology and function, were collected consistently from a young adult rat pancreas.