2011 - IPITA - Prague


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Parallel session 4 – Open oral presentations Topic: Islet transplantation: Technical aspects

4.9 - Development of a PFC-enriched fibrin clot system for islet culture

Presenter: E., Maillard, Oxford, United Kingdom
Authors: E. Maillard, M.T. Juszczak, A. Clark, S.J. Hughes, D.W.R. Gray, P.R.V. Johnson


Development of a PFC-enriched fibrin clot system for islet culture

 

E. Maillard, M.T. Juszczak, A. Clark, S.J. Hughes, D.W.R. Gray, P.R.V. Johnson
University of Oxford, Nuffield Department of Surgical Sciences, Oxford, U.K.

Background: Whilst preservation of viability and secretory function has been demonstrated after culturing islets in fibrin, limited oxygen diffusion may limit its clinical applicability. The aim of this study was to determine whether the combination of fibrin and perfluorodecalin could create a suitable microenvironment for human islets in culture.

Material and methods: Islets from 8 human pancreases were cultured immediately after isolation in either control/standard conditions or in a fibrin gel (10mg/mL) supplemented with 10% of raw or emulsified perfluorodecalin. After 24h islet viability (FDA/EB, caspase-3), function (modified GSIS index, release of insulin and pro-insulin) and expression of hypoxia markers (HIF-1α translocation into nucleus and expression of VEGF) were assessed.

Results: Islet viability was similar in all conditions tested, however presence of fibrin resulted in about 50% decrease in caspase-3 activation as compared with standard culture (0.60±0.12ng vs. 1.17±0.10ng of activated caspase-3/μg of protein; p<0.05). A significant reduction in insulin and proinsulin release was observed when islets were cultured in fibrin gel as compared with standard conditions (6.65±0.87pg vs. 19.44±4.33pg pro-insulin/μg protein; p<0.05), irrespective of presence of PDC. Islets cultured in standard conditions and in fibrin alone had impaired stimulation indices (0.98±0.17 and 1.18±10.33 respectively). A significant, 3-fold increase in stimulation index was observed in islets cultured in the presence of fibrin gel supplemented with emulsified but not raw PDC (3.01±0.84 vs. 1.58±0.45; p<0.05). Emulsified PDC in fibrin gels decreased and raw PDC increased translocation of HIF-1α and VEGF secretion compared with fibrin alone (1.01±0.23 vs. 1.94±0.80 vs. 1.45±0.41 and 14.62±3.36 vs. 32.60±3.36pg vs. 20.91±3.55pg VEGF/μg protein, respectively).

Conclusions: Supplementation of fibrin gels with emulsified perfluorodecalin reduces islet hypoxia as indicated by HIF-1a translocation and release of VEGF and improves islet secretory function. However, the formulation of the PDC plays a crucial role for its ability to prevent hypoxia.

Parallel session 9 - Oral presentations - Topic: Pancreas transplantation: Long-term function and rejection


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