Young porcine endocrine pancreatic islets cultured in fibrin and alginate gels show improved resistance towards human monocytes

Abstract

AimTo investigate the protective function of alginate and fibrin gels used to embed porcine endocrine pancreatic islets towards human monocytes. MethodsGroups of 200 islet equivalents from young pigs were embedded in either a fibrin or in an alginate gel, and as a control seeded in tissue culture polystyrene (TCPS) well plates. The islet cultures were incubated with 2×105 human monocytes for 24h. In addition, both islets and monocytes were separately cultured in TCPS, fibrin and alginate. Islet morphology, viability and function were investigated as well as the secretion of cytokines TNFα, IL-6, and IL-1β. ResultsWhen freely-floating in TCPS, non-encapsulated islets were surrounded by monocytes and started to disperse after 24h. In fibrin, monocytes could be found in close proximity to embedded islets, indicating monocyte migration through the gel. In contrast, after 24h, few monocytes were found close to islets in alginate. Immunofluorescence staining and manual counting showed that integrin expression was higher in fibrin-embedded islet cultures. A TUNEL assay revealed elevated numbers of apoptotic cells for islets in TCPS wells compared to fibrin and alginate cultures. Insulin secretion was higher with islets embedded in fibrin and alginate when compared to non-encapsulated islets. TNFα, IL-6 and IL-1β were found in high concentrations in the media of co-cultures and monocyte mono-culture in fibrin. ConclusionBoth alginate and fibrin provide key structural support and offer some protection for the islets towards human monocytes. Fibrin itself triggers the cytokine secretion from monocytes.