Abstract
Transplantation of pancreatic islet cells derived from human pluripotent stem cells is a promising treatment for diabetes. Despite progress in the generation of stem-cell-derived islets (SC-islets), no detailed characterization of their functional properties has been conducted. Here, we generated functionally mature SC-islets using an optimized protocol and benchmarked them comprehensively against primary adult islets. Biphasic glucose-stimulated insulin secretion developed during in vitro maturation, associated with cytoarchitectural reorganization and the increasing presence of alpha cells. Electrophysiology, signaling and exocytosis of SC-islets were similar to those of adult islets. Glucose-responsive insulin secretion was achieved despite differences in glycolytic and mitochondrial glucose metabolism. Single-cell transcriptomics of SC-islets in vitro and throughout 6 months of engraftment in mice revealed a continuous maturation trajectory culminating in a transcriptional landscape closely resembling that of primary islets. Our thorough evaluation of SC-islet maturation highlights their advanced degree of functionality and supports their use in further efforts to understand and combat diabetes.
Highlights
The generation of functional pancreatic beta cells from human pluripotent stem cells is a main goal of stem cell research, aiming to provide a renewable and consistent source of cells for the treatment of diabetes
At S7w0, SC-islets contained ≈40% insulin-positive monohormonal cells (INS+)—a proportion that remained relatively stable until S7w6 (Fig. 1b,c and Supplementary Fig. 1b,c)
Through indepth functional assays, cell physiology analyses, metabolic tracing experiments and single-cell RNA (scRNA) transcriptomic data throughout 6 weeks of in vitro maturation and 6 months of mouse engraftment, we show the temporal acquisition of metabolic programs and gene regulatory changes that contribute to beta cell functional maturation (Fig. 6k)
Summary
The generation of functional pancreatic beta cells from human pluripotent stem cells (hPSCs) is a main goal of stem cell research, aiming to provide a renewable and consistent source of cells for the treatment of diabetes. Individual studies have reported particular transcriptomic[7,10], functional[3,8,9,11] and metabolic[2,12] aspects of SC-islets Studies integrating these aspects with detailed analyses of stimulus-secretion coupling and exocytosis machinery of functional SC-islets have been lacking. We conducted detailed functional and physiological characterization of the SC-islets, supported by targeted metabolite tracing studies together with single-cell transcriptomic profiling of differentiating endocrine cell populations. This multipronged approach was conducted both during the timecourse of SC-islet maturation in vitro and after in vivo engraftment. Our integrated analyses show that a high level of beta cell functionality is achieved in vitro even if specific metabolic and transcriptomic differences persist between SC-islet beta cells and primary beta cells
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