Will be more effective in smaller endocrine 871361-88-5 web aggregates enhancing their survival and function until the re-establishment of the islet vasculature. Small islet aggregates have previously been shown to be superior to large intact islets as graft material in diabetic mice, with improved transplantation outcomes being associated with reduced hypoxia-related necrosis in the small islet aggregates [36]. Importantly, this benefit of small islet aggregates over large intact islets was demonstrated using encapsulated islets which do not revascularise in vivo, so the improved islet function was independent of any influence on islet revascularisation. Graft revascularisation is obviously important for subsequent function and inadequate revascularisation of transplanted islets at a number of implantation sites is associated with deleterious outcomes [37?0], whereas improvements in graft revascularisation are associated with improved islet function and long-term survival [41?3]. Our results demonstrate that maintaining individual islets at the graft site resulted in a significant enhancement of revascularisation, consistent with a previousFigure 7. Efficacy of matrigel-implanted islets in vivo. A. Blood glucose concentrations of mice transplanted with pelleted islets alone (continuous line) or islets dispersed in matrigel (dashed line), beneath the kidney capsule, *p,0.05, Two-Way RM ANOVA with Bonferroni post hoc test, n = 7-8. B. Percentage of mice remaining diabetic (blood glucose concentration .11.1 mmol/l) after transplantation as in A, p = 0.02 Kaplan eier, n = 7-8. doi:10.1371/journal.pone.0057844.greport of superior revascularisation of small, compared to larger islets 
[44]. Similarly, in our previous study where we cotransplanted islets with MSCs, the resultant smaller endocrine aggregates had an enhanced vascular density compared to that of the large endocrine masses formed in mice implanted with islets alone [6]. Intra-islet interactions are known to be important for normal islet function [45,46] and disruption of islet architecture is associated with impaired secretory responses to a range of physiological stimuli. Maintaining anatomically correct islet architecture may therefore further enhance graft function by facilitating the numerous interactions between islet cells [47] that are required for normal insulin secretion [45,46]. Our observations using the renal subcapsular graft site are in accordance with recent studies of intramuscular islet transplantation, in which islets grafted as clusters developed central fibrosis [48], whereas transplanting the islets in a `pearls-on-a-string’ configuration, such that they are engrafted essentially as single islets, was associated with improved transplantation outcomesMaintenance of Islet Morphology[49]. This suggests that the beneficial impact of maintaining islet anatomy during transplantation is not graft P7C3 cost site-specific. In conclusion, there is mounting evidence that the current intraportal route for clinical islet transplantation places the grafts into a hostile microenvironment and confers multiple and perhaps avoidable stresses upon the transplanted islets [3], so efforts are being made to identify alternative optimal implantation sites for islets. The current study suggests that preventing the fusion of islets at extrahepatic sites represents an important strategy for promoting islet engraftment, which may contribute to achieving routine single donor islet transplantation [2,50], thereby.Will be more effective in smaller endocrine aggregates enhancing their survival and function until the re-establishment of the islet vasculature. Small islet aggregates have previously been shown to be superior to large intact islets as graft material in diabetic mice, with improved transplantation outcomes being associated with reduced hypoxia-related necrosis in the small islet aggregates [36]. Importantly, this benefit of small islet aggregates over large intact islets was demonstrated using encapsulated islets which do not revascularise in vivo, so the improved islet function was independent of any influence on islet revascularisation. Graft revascularisation is obviously important for subsequent function and inadequate revascularisation of transplanted islets at a number of implantation sites is associated with deleterious outcomes [37?0], whereas improvements in graft revascularisation are associated with improved islet function and long-term survival [41?3]. Our results demonstrate that maintaining individual islets at the graft site resulted in a significant enhancement of revascularisation, consistent with a previousFigure 7. Efficacy of matrigel-implanted islets in vivo. A. Blood glucose concentrations of mice transplanted with pelleted islets alone (continuous line) or islets dispersed in matrigel (dashed line), beneath the kidney capsule, *p,0.05, Two-Way RM ANOVA with Bonferroni post hoc test, n = 7-8. B. Percentage of mice remaining diabetic (blood glucose concentration .11.1 mmol/l) after transplantation as in A, p = 0.02 Kaplan eier, n 
= 7-8. doi:10.1371/journal.pone.0057844.greport of superior revascularisation of small, compared to larger islets [44]. Similarly, in our previous study where we cotransplanted islets with MSCs, the resultant smaller endocrine aggregates had an enhanced vascular density compared to that of the large endocrine masses formed in mice implanted with islets alone [6]. Intra-islet interactions are known to be important for normal islet function [45,46] and disruption of islet architecture is associated with impaired secretory responses to a range of physiological stimuli. Maintaining anatomically correct islet architecture may therefore further enhance graft function by facilitating the numerous interactions between islet cells [47] that are required for normal insulin secretion [45,46]. Our observations using the renal subcapsular graft site are in accordance with recent studies of intramuscular islet transplantation, in which islets grafted as clusters developed central fibrosis [48], whereas transplanting the islets in a `pearls-on-a-string’ configuration, such that they are engrafted essentially as single islets, was associated with improved transplantation outcomesMaintenance of Islet Morphology[49]. This suggests that the beneficial impact of maintaining islet anatomy during transplantation is not graft site-specific. In conclusion, there is mounting evidence that the current intraportal route for clinical islet transplantation places the grafts into a hostile microenvironment and confers multiple and perhaps avoidable stresses upon the transplanted islets [3], so efforts are being made to identify alternative optimal implantation sites for islets. The current study suggests that preventing the fusion of islets at extrahepatic sites represents an important strategy for promoting islet engraftment, which may contribute to achieving routine single donor islet transplantation [2,50], thereby.
