Abstract
An automated method for the isolation of human pancreatic islets is described. The procedure meets the following requirements: 1) minimal traumatic action on the islets, 2) continuous digestion in which the islets that are progressively liberated can be saved from further enzymatic action, 3) minimal human intervention in the digestion process, and 4) high yield and purity of the isolated islets. After purification of Ficoll gradients, an average of 2279 islets/g pancreas was obtained, with an average purity of 79% islets. The average volume and average insulin content of the final islet preparation were 348 mm3 and 93.4 U, respectively. The islets were morphologically intact with a normal degree of beta-cell granulation, responded to glucose stimulation with a fivefold increase of insulin secretion over basal levels, and produced normoglycemia after transplantation into diabetic mice. The procedure is being used in the second phase of clinical trials of islet transplantation in patients with insulin-dependent diabetes mellitus.
