Donor Islet Endothelial Cells Participate in Formation of Functional Vessels Within Pancreatic Islet Grafts

Nyqvist, Daniel; Köhler, Martin; Wahlstedt, Helene; Berggren, Per Olof

doi:10.2337/diabetes.54.8.2287

Cited by 157 publications

(188 citation statements)

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“…2a-e). These data extend previous reports based on two-photon microscopy showing that, after an initial phase of vascularisation, syngeneic islets transplanted into the ACE are sustained and function over several months [20,25].…”

Section: Resultssupporting

confidence: 78%

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Longitudinal three-dimensional visualisation of autoimmune diabetes by functional optical coherence imaging

et al. 2015

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Aims/hypothesis It is generally accepted that structural and functional quantitative imaging of individual islets would be beneficial to elucidate the pathogenesis of type 1 diabetes. We here introduce functional optical coherence imaging (FOCI) for fast, label-free monitoring of beta cell destruction and associated alterations of islet vascularisation. Methods NOD mouse and human islets transplanted into the anterior chamber of the eye (ACE) were imaged with FOCI, in which the optical contrast of FOCI is based on intrinsic variations of the index of refraction resulting in a faster tomographic acquisition. In addition, the phase sensitivity allows simultaneous label-free acquisition of vascularisation. Results We demonstrate that FOCI allows longitudinal quantification of progressive autoimmune insulitis, including the three-dimensional quantification of beta cell volume, inflammation and vascularisation. The substantially increased backscattering of islets is dominated by the insulin-zinc nanocrystals in the beta cell granules. This translates into a high specificity for the functional beta cell volume of islets. Applying FOCI to a spontaneous mouse model of type 1 diabetes, we quantify the modifications of the pancreatic microvasculature accompanying the progression of diabetes and reveal a strong correlation between increasing insulitis and density of the vascular network of the islet. Conclusions/interpretation FOCI provides a novel imaging technique for investigating functional and structural diabetes-induced alterations of the islets. The label-free detection of beta cell volume and infiltration together with vascularisation offers a unique extension to study ACE-transplanted Diabetologia (2016) 59:550-559 DOI 10.1007 Corinne Berclaz, Anja Schmidt-Christensen and Daniel Szlag contributed equally to this study. Electronic supplementary materialThe online version of this article (doi:10.1007/s00125-015-3819-x) contains peer-reviewed but unedited supplementary material, which is available to authorised users.

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Section: Resultssupporting

confidence: 78%

“…Pancreatic islet isolation, culture and ACE transplantation Mouse pancreatic islets were isolated according to the protocol previously described [25]. Between 10 and 50 cultured islets were transplanted per eye, as previously described [21].…”

Section: Methodsmentioning

confidence: 99%

Longitudinal three-dimensional visualisation of autoimmune diabetes by functional optical coherence imaging

et al. 2015

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“…Islet incubation in ECCM Twenty islets were precultured for 4-7 days in CM, and subsequently cultured in 1 ml of control CM or ECCM in a 24 well culture dish for 24 h. The reason for the pre-culture period was to minimise remnant islet endothelial cells, as studies have shown that~9 0% of the intra-islet endothelial cells disappear after 4 days in culture [6,19].…”

Section: Isolation Of Islet Endothelial Cellsmentioning

confidence: 99%

Endothelial cell signalling supports pancreatic beta cell function in the rat

et al. 2009

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Aims/hypothesis The proximity of endothelial cells and beta cells in islets by necessity means that they are exposed to each other's products. Whereas islet endothelial cells require signals from beta cells to function properly, endothelin-1, thrombospondin-1 and laminins, among others, have been identified as endothelial-derived molecules, although their full effects on beta cells have not been explored. We tested the hypothesis that islet endothelialderived products affect beta cell function. Methods Endothelial cells from rat islets were proliferated and purified. Endothelium-conditioned culture medium (ECCM) was obtained by maintaining the endothelial cells in culture medium. Islet function was evaluated following exposure of cultured islets to standard culture medium or ECCM. Changes in mRNA levels for key beta cell metabolic enzymes were also measured in islets after ECCM exposure.Results Glucose-stimulated insulin release and islet insulin content were markedly enhanced by exposure to ECCM. This was at least partly explained by improved mitochondrial function, as assessed by glucose oxidation and an upregulation of the mitochondrial gene for glycerol-3-phosphate dehydrogenase (mGpdh [also known as Gpd2]), combined with upregulation of the rate-limiting enzyme in the glycolysis, glucokinase, in the islets. The intracellular degradation of insulin was also decreased in the islets. Islet endothelial cells produced laminins, and the positive effects of islet endothelial cells were prevented by addition of a neutralising antibody to the β1-chain of laminin. Addition of exogenous laminin stimulated islet function. Conclusions/interpretation This study provides proof of principle that endothelial cells can affect the function of beta cells in their vicinity and that this is at least partially mediated by laminins.

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“…We isolated and cultured pancreatic islets as previously described 26,29 . For transplantation, we transferred 30-300 islets from culture media to sterile PBS and aspirated them into a blunt 27-gauge eye cannula connected to a 1-ml Hamilton syringe (Hamilton) via 0.4-mm polythene tubing (Portex Limited).…”

Section: Transplantation Of Pancreatic Islets To the Anterior Chambermentioning

confidence: 99%

Noninvasive in vivo imaging of pancreatic islet cell biology

Speier

Nyqvist

Cabrera

et al. 2008

Nat Med

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Advanced imaging techniques have become a valuable tool in the study of complex biological processes at the cellular level in biomedical research. Here, we introduce a new technical platform for noninvasive in vivo fluorescence imaging of pancreatic islets using the anterior chamber of the eye as a natural body window. Islets transplanted into the mouse eye engrafted on the iris, became vascularized, retained cellular composition, responded to stimulation and reverted diabetes. Laserscanning microscopy allowed repetitive in vivo imaging of islet vascularization, beta cell function and death at cellular resolution. Our results thus establish the basis for noninvasive in vivo investigations of complex cellular processes, like beta cell stimulus-response coupling, which can be performed longitudinally under both physiological and pathological conditions. Adequate release of insulin by pancreatic beta cells in response to changing blood glucose levels is a vital requirement for maintaining glucose homeostasis. Failure to do so is one of the major causes of type 2 diabetes mellitus, the most common metabolic disorder in humans 1 . Under physiological conditions, insulin release is regulated by the complex interplay between glucose and a plethora of additional factors-for example, nutrients, autocrine-paracrine signaling and the continuous input from hormones and neurotransmitters 2 . Beta cells, together with other pancreatic endocrine cell types, are situated within the endocrine pancreas, that is, the islets of Langerhans, which are densely vascularized 3 and abundantly innervated 4 . Pancreatic islets, constituting 1%-2% of the pancreatic volume, are difficult to access for in vivo monitoring because they are deeply embedded and scattered in the exocrine tissue of the pancreas 5 . As a consequence, the majority of functional beta cell studies have so far been conducted in vitro on isolated islets or beta cells. Isolated islets 6 , and especially pancreatic slices 7 , allow functional studies of Author Contributions: S.S., D.N. and A.C. developed the experimental transplantation platform. S.S., D.N., O.C., J.Y., R.D.M., A.P., T.M., M.K., B.L. and A.C. did the experiments. J.W. was responsible for generating the transgenic mice. C.R. was involved in designing the transplantation protocols and writing the manuscript. S.S., D.N., I.B.L. and P.-O.B. were responsible for designing the overall experimental plan and writing the manuscript. P.-O.B. was the originator of the idea of using the anterior chamber of the eye for noninvasive in vivo imaging of pancreatic islet cell biology Reprints and permissions information is available online at http://npg.nature.com/reprintsandpermissions Note: Supplementary information is available on the Nature Medicine website. Laser-scanning microscopy (LSM) of isolated islets and cell preparations has been successfully applied for imaging multiple signaling pathways in the beta cell 6 . However, intravital applications of LSM for studies of beta cell physiology have not been repo...

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Donor Islet Endothelial Cells Participate in Formation of Functional Vessels Within Pancreatic Islet Grafts

Cited by 157 publications

References 23 publications

Longitudinal three-dimensional visualisation of autoimmune diabetes by functional optical coherence imaging

Longitudinal three-dimensional visualisation of autoimmune diabetes by functional optical coherence imaging

Endothelial cell signalling supports pancreatic beta cell function in the rat

Noninvasive in vivo imaging of pancreatic islet cell biology

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