Young porcine endocrine pancreatic islets cultured in fibrin show improved resistance toward hydrogen peroxide

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Abstract

Aim:

To study the protective effect of a fibrin scaffold toward embedded young porcine endocrine pancreatic islets from hydrogen peroxide within the context of islet encapsulation in transplantation.

Methods:

After isolation and in vitro maturation, groups of 200 young porcine islet equivalents (IEQ) were embedded in a 200 μL fibrin gel and exposed to 2 concentrations (10 and 100 μM) of hydrogen peroxide (H2O2) to investigate the ability of fibrin to protect islets against apoptotic stimuli. As a control, young porcine islets were seeded in tissue culture polystyrene (TCPS) well plates and exposed to the same H2O2 concentrations. Islet integrity, viability and function were then investigated.

Results:

Morphologically, the integrity of islets embedded in fibrin gels was better preserved compared with that of islets cultured in TCPS plates, when exposed to H2O2. Immunofluorescence staining showed that insulin and glucagon expression was higher in islets cultured in fibrin. Overall, H2O2 incubation led to decreased insulin and glucagon expression. A TUNEL assay revealed elevated numbers of apoptotic cells for islets cultured in TCPS plates when compared with those embedded in fibrin. Islets cultured in TCPS plates and exposed to H2O2 had diminished ability to secrete insulin in response to glucose stimulation, whereas islets embedded in fibrin maintained their glucose responsiveness. Insulin trapped in fibrin was extracted and quantified, revealing insulin in the extract.

Conclusions/Interpretation:

Fibrin has a protective effect on young porcine endocrine pancreatic islets exposed to hydrogen peroxide.

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