Islets in the body are never flat: transitioning from two-dimensional (2D) monolayer culture to three-dimensional (3D) spheroid for better efficiency in the generation of functional hPSC-derived pancreatic β cells in vitro

Abstract

Diabetes mellitus (DM), currently affecting more than 537 million people worldwide is a chronic disease characterized by impaired glucose metabolism resulting from a defect in insulin secretion, action, or both due to the loss or dysfunction of pancreatic β cells. Since cadaveric islet transplantation using Edmonton protocol has served as an effective intervention to restore normoglycaemia in T1D patients for months, stem cell-derived β cells have been explored for cell replacement therapy for diabetes. Thus, great effort has been concentrated by scientists on developing in vitro differentiation protocols to realize the therapeutic potential of hPSC-derived β cells. However, most of the 2D traditional monolayer culture could mainly generate insulin-producing β cells with immature phenotype. In the body, pancreatic islets are 3D cell arrangements with complex cell–cell and cell–ECM interactions. Therefore, it is important to consider the spatial organization of the cell in the culture environment. More recently, 3D cell culture platforms have emerged as powerful tools with huge translational potential, particularly for stem cell research. 3D protocols provide a better model to recapitulate not only the in vivo morphology, but also the cell connectivity, polarity, and gene expression mimicking more physiologically the in vivo cell niche. Therefore, the 3D culture constitutes a more relevant model that may help to fill the gap between in vitro and in vivo models. Interestingly, most of the 2D planar methodologies that successfully generated functional hPSC-derived β cells have switched to a 3D arrangement of cells from pancreatic progenitor stage either as suspension clusters or as aggregates, suggesting the effect of 3D on β cell functionality. In this review we highlight the role of dimensionality (2D vs 3D) on the differentiation efficiency for generation of hPSC-derived insulin-producing β cells in vitro. Consequently, how transitioning from 2D monolayer culture to 3D spheroid would provide a better model for an efficient generation of fully functional hPSC-derived β cells mimicking in vivo islet niche for diabetes therapy or drug screening.

Exploring the functional, protective, and transcriptomic effects of GIP on cytokine-exposed human pancreatic islets and EndoC-βH5 cells

“…The copyright holder for this preprint this version posted June 18, 2024. ; https://doi.org/10.1101/2024.06.17.599254 doi: bioRxiv preprint 12 cells are absent [50,51]. This is further supported by the fact that we did not observe GIP-induced insulin secretion at low glucose in human islets in both this and a previous study [12].…”

“…As recently reported for the newly established non-proliferative human beta-cell line EndoC-βH5 [26], we found that GIP acutely augmented insulin secretion at high glucose (20 mM) over a range of supraphysiological doses and modestly increased insulin secretion at low (2 mM) glucose. The increase in insulin secretion at low glucose may reflect that the tight regulation of beta-cell function is lost in 2D culture with a single islet cell type where paracrine signals from the delta and alpha 12 cells are absent [50,51]. This is further supported by the fact that we did not observe GIP-induced insulin secretion at low glucose in human islets in both this and a previous study [12].…”