A simple, reliable method for high-throughput screening for diabetes drugs using 3D beta-cell spheroids

摘要

Early screens for new diabetes drugs rely on monolayers of beta-cells, which are known to be poor predictors of the in vivo response. Previously, we developed a method to create uniform islet spheroids from freshly-dispersed human donor tissue for drug screening. While the human engineered islets worked well to reduce donor-to-donor variability, it is difficult and expensive to obtain sufficient high-quality human islets for drug testing. Thus, this study utilized a genetically-modified beta-cell culture line (INS-1832/13) in 2D and as 3D spheroids and compared the results to human islet tissue formed into spheroids using a high-throughput 384-well format. In response to increasing concentrations of glucose, all 3 groups increased insulin release, but the cultured beta-cells (2D and 3D) were more sensitive to glucose (EC50 5.85 mM for 2D beta-cells, 16.24 mM for 3D beta-cell spheroids) than the human islet spheroids (EC50 53.69 mM). The order of responses to glybenclamide was human spheroids > 3D beta-cell culture > 2D beta-cell culture. In response to caffeine, the beta-cells in 2D or 3D were more responsive compared to the human islet spheroids (EC50 0.39 and 0.31 mM for 2D and 3D beta-cells respectively). When exposed to inhibitors of insulin secretion (nifedipine and diazoxide), the responses were more similar between groups. Z' calculations, indicative of assay quality, determined that the 3D beta-cell spheroids reached the criteria of an excellent to ideal drug screen assay more consistently than the other test models. In conclusion, 3D beta-cell spheroids from a cultured cell line can be used in HTS assays that, according to reference drugs tested here, are sensitive and predictive of the in vivo response. (C) 2016 The Authors. Published by Elsevier Inc.