Effective endothelial cell and human pluripotent stem cell interactions generate functional insulin-producing beta cells

Abstract

Aims/hypothesisEndothelial cells (ECs) play an essential role in pancreatic organogenesis. We hypothesise that effective in vitro interactions between human microvascular endothelial cells (HMECs) and human pluripotent stem cells (hPSCs) results in the generation of functional pancreatic beta cells.MethodsEmbryoid bodies (EBs) derived from hPSCs were cultured alone (controls) or with ECs in collagen gels. Subsequently, cells were analysed for pancreatic beta cell markers, and then isolated and expanded. Insulin secretion in response to glucose was evaluated in vitro by static and dynamic (perifusion) assays, and in vivo by EB transplantation into immunodeficient mice.ResultsCo-cultured EBs had a higher expression of mature beta cells markers and enhanced insulin secretion in vitro, compared with controls. In mice, transplanted EBs had higher levels of human C-peptide secretion with a significant reduction in hyperglycaemia after the selective destruction of native pancreatic beta cells. In addition, there was significant in vitro upregulation of bone morphogenetic proteins 2 and 4 (BMP-2, 4) in co-cultured cells, compared with controls.Conclusions/interpretationECs provide essential signalling in vitro, such as activation of the BMP pathway, for derivation of functional insulin-producing beta cells from hPSCs.Electronic supplementary materialThe online version of this article (doi:10.1007/s00125-016-4078-1) contains peer-reviewed but unedited supplementary material, which is available to authorised users.

Design Considerations for Macroencapsulation Devices for Stem Cell Derived Islets for the Treatment of Type 1 Diabetes

“…In particular, this has been done with endothelial cells that, while normally present in primary islets, [20] are not produced with the differentiation protocols that generate SC-islets. [40][41][42][43] However, while the theoretical inclusion of endothelial cells should add noticeable improvements in SC-islets, to-date the published protocols have not improved upon the results of publications without their inclusion. [34,44] Preventing the immune rejection of these cells after transplantation is a current major hurdle to clinical translation.…”

The Feasibility and Applicability of Stem Cell Therapy for the Cure of Type 1 Diabetes

“…When MSCs from the human pancreas are cultured with dimethyloxallyl glycine and TNF-α, nuclear factor erythroid 2-related factor 2 (NRF2) and VEGF are upregulated; these factors may promote graft revascularization and graft survival. Endothelial cells also act effectively on β-cells maturation and survival [ 79 ]. Native β-cells exhibit polarity, with an apical and basolateral membrane and a greater density of insulin vesicles in the basal region, close to the endothelial cells.…”

Harnessing Proliferation for the Expansion of Stem Cell-Derived Pancreatic Cells: Advantages and Limitations

“…The end goal of stem cell-derived b-like cell research is to develop a transplantable system of cells that fully replicate b-cell function. Ideal tactics to achieve this would be to optimize current differentiation protocols, such as through manual purification of mature populations or treatments that increase final b-like cells (10,12,41,52,60,61), and to improve in vivo site support for transplanted b-like cells (62)(63)(64). Despite the limitations listed above, expanding b-like cells or their progenitors in vitro still presents a promising strategy to further support these approaches.…”