Advances toward engineering functionally mature human pluripotent stem cell-derived β cells

Human stem cell-derived β (SC-β) cells have the potential to revolutionize diabetes treatment through disease modeling, drug screening, and cellular therapy. SC-β cells are likely to represent an early clinical translation of differentiated human pluripotent stem cells (hPSC). In 2014, two groups generated the firs

SIX2 regulates human β cell differentiation from stem cells and functional maturation in vitro

Generation of insulin-secreting β cells in vitro is a promising approach for diabetes cell therapy. Human embryonic stem cells (hESCs) and human induced pluripotent stem cells (hiPSCs) are differentiated to β cells (SC-β cells) and mature to undergo glucose-stimulated insulin secretion, but molecular regulation of this defining β cell phenotype is unknown. Here, we show that maturation of SC-β cells is regulated by the transcription factor SIX2. Knockdown (KD) or knockout (KO) of SIX2 in SC-β cells drastically limits glucose-stimulated insulin secretion in both static and dynamic assays, along with the upstream processes of cytoplasmic calcium flux and mitochondrial respiration. Furthermore, SIX2 regulates the expression of genes associated with these key β cell processes, and its expression is restricted to endocrine cells. Our results demonstrate that expression of SIX2 influences the generation of human SC-β cells in vitro

Acquisition of Dynamic Function in Human Stem Cell-Derived β Cells

Summary: Recent advances in human pluripotent stem cell (hPSC) differentiation protocols have generated insulin-producing cells resembling pancreatic β cells. While these stem cell-derived β (SC-β) cells are capable of undergoing glucose-stimulated insulin secretion (GSIS), insulin secretion per cell remains low compared with islets and cells lack dynamic insulin release. Herein, we report a differentiation strategy focused on modulating transforming growth factor β (TGF-β) signaling, controlling cellular cluster size, and using an enriched serum-free media to generate SC-β cells that express β cell markers and undergo GSIS with first- and second-phase dynamic insulin secretion. Transplantation of these cells into mice greatly improves glucose tolerance. These results reveal that specific time frames for inhibiting and permitting TGF-β signaling are required during SC-β cell differentiation to achieve dynamic function. The capacity of these cells to undergo GSIS with dynamic insulin release makes them a promising cell source for diabetes cellular therapy. : In this study, Millman and colleagues report a differentiation strategy to generate β-like cells from human pluripotent stem cells with islet-like dynamic insulin release that rapidly reverses diabetes in mice. The authors elucidate that stage-specific control of TGF-β signaling during endocrine induction and maturation to be critical for robust function. Keywords: human embryonic stem cells, human induced pluripotent stem cells, diabetes, differentiation, glucose-stimulated insulin secretion, transplantation, cell therapy, β cells, pancreas, islet