Ex Vivo Expansion and Differentiation of Human and Mouse Fetal Pancreatic Progenitors Are Modulated by Epidermal Growth Factor

Abstract

A comparative analysis of mouse and human pancreatic development may reveal common mechanisms that control key steps as organ morphogenesis and cell proliferation and differentiation. More specifically, understanding beta cell development remains an issue, despite recent progress related to their generation from human embryonic and induced pluripotent stem cells. In this study, we use an integrated approach, including prospective isolation, organ culture, and characterization of intermediate stages, and report that cells from human and mouse fetal pancreas can be expanded in the long term and give rise to hollow duct-like structures in 3D cultures. The expanded cells express a combination of markers (E-cadherin, PDX1, NKX6-1, SOX9, and HNF1β) that reveals pancreatic progenitor identity. Proliferation of embryonic progenitors was stimulated by the Wnt agonist R-spondin1 (RSPO1), FGF10, and EGF. This combination of growth factors allowed maintaining human fetal pancreatic progenitors in culture for many passages, a finding not reported previously. Importantly, in the absence of EGF, proliferation was reduced, while endocrine differentiation was significantly enhanced. We conclude that modulation of EGF signaling affects in vitro expansion and differentiation of progenitors from embryonic pancreas of both mice and man.

Human pancreas development

“…These data align with those from mouse (Rodríguez-Seguel et al, 2013) and, most recently, with results from an exciting new methodology for long-term culture of human pancreatic progenitors showing positive effects of WNT, FGF10 and epidermal growth factor (EGF) signaling on cell proliferation (Bonfanti et al, 2015). Alongside this pro-proliferative maintenance of progenitor status, EGF also inhibited endocrine differentiation (Bonfanti et al, 2015). To date, the role of Notch and retinoic acid signaling in promoting progenitor cell proliferation, or bone morphogenetic protein (BMP) signaling in inhibiting it, has not been directly analyzed during human embryogenesis, although these factors are known to be important in the mouse (Jørgensen et al, 2007;Pan and Wright, 2011).…”

“…By comparative transcriptomics, gene ontology analysis highlighted enrichment for components of WNT signaling in human pancreatic progenitors at CS16-CS18, prior to significant acinar cell differentiation and before noticeable endocrine differentiation (Cebola et al, 2015). These data align with those from mouse (Rodríguez-Seguel et al, 2013) and, most recently, with results from an exciting new methodology for long-term culture of human pancreatic progenitors showing positive effects of WNT, FGF10 and epidermal growth factor (EGF) signaling on cell proliferation (Bonfanti et al, 2015). Alongside this pro-proliferative maintenance of progenitor status, EGF also inhibited endocrine differentiation (Bonfanti et al, 2015).…”

β-cell replacement sources for type 1 diabetes: a focus on pancreatic ductal cells

“…These results were confirmed by Cavelti-Weder and colleagues, who showed extensive periods (up to 13 months) of euglycemia in two of the TF-transduced diabetic animals . Similarly, a cytokine-based protocol [containing epidermal growth factor (EGF) and ciliary neurotrophic factor (CNTF)] was developed to circumvent the use of adenoviruses for acinar-to-β transdifferentiation [Bonfanti et al 2015]. In this study, alloxan-induced diabetic mice received EGF and CNTF via intraperitoneal osmotic pumps that allowed 65% of animals to increase their levels of serum insulin and normalize their glycemia within 5 days.…”

Diabetes through a 3D lens: organoid models

“…Tissues controlling glucose homeostasis Pancreas The first pancreatic organoids were generated from mouse and human embryonic pancreatic cells by Anne Grapin-Botton's group [43,44]. These organoids produced progenitor-biased hollow spheres that could be differentiated into branched structures containing acinar, ductal and endocrine lineages.…”