Enzymatic digestion of donor pancreas is a vital step in islet isolation process. Recently, human and large mammalian islet isolation has been improved by Liberase. However, there are no data to show the improvement of rat islet isolation with Liberase. We hypothesized that commercially available Liberase has variable activities from batch to batch and that a short culture period might improve the function of isolated islets in vivo. We therefore isolated islets with Liberase and cultured them for a short period before transplantation to reverse diabetes in a syngenic rat model.Methods:
Islets of high inbred Lewis rats were isolated with Liberase, purified by discontinuous density gradients, hand-picked, and cultured at 37°C for 24 hours. The same batch of Liberase was used for all experiments in this study. Freshly isolated and cultured islets were implanted intraportally into rats rendered diabetic by streptozotocin.Results:
In this study, we were not able to reverse diabetes by syngenic transplantation of freshly isolated islets from 2 donors with Liberase in the rat model. Surprisingly, for the first time, we successfully achieved normoglycemia for more than 100 days in the diabetic rats by syngenic transplantation of the cultured islets isolated with Liberase. These normoglycemic diabetic rats showed normal glucose tolerance curves. Histologic examination of the livers of the islet recipients revealed intact islets, with numerous well-granulated insulin-containing cells and only few glucagon-expressing cells. In islets of the recipients' pancreata, the remaining native islets consisted mainly of glucagon-expressing cells, with few insulin-expressing cells in the center.Conclusions:
We conclude that Liberase isolation followed by a short culture period may be a good substitute for collagenase in rats. Islet culture before transplantation can promote the success of single-donor-single-recipient islet transplantation to reverse diabetes in the rat model.