Pseudoislets as primary islet replacements for research

Persaud, Shanta J.; Arden, Catherine; Bergsten, Peter; Bone, Adrian J.; Brown, James E.; Dunmore, Simon; Harrison, Moira; Hauge-Evans, Astrid C.; Kelly, Catriona; King, Aileen; Maffucci, Tania; Marriott, Claire; McClenaghan, Neville H.; Morgan, Noel G.; Reers, Christina; Russell, Mark A.; Turner, Mark D.; Willoughby, Emma; Younis, MustafaY.G.; Zhi, Zheng‐liang; Jones, Peter M.

doi:10.4161/isl.2.4.12557

Cited by 16 publications

(6 citation statements)

References 19 publications

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“…PI formation improves β cell function overtime [4], [5], [6], [7], [8], [9]. In our hands, MS1-induced PIs included an increased proportion of insulin-positive cells when compared with βTC3 monolayers.…”

Section: Discussionsupporting

confidence: 48%

“…In contrast to monolayer β-cells, PIs represent a more native structure for β cell cultures capable of improving β cell function. This improvement is partially attributed to enhanced cell-cell contact leading to increased glucose sensing, insulin production, and insulin release [4], [5], [6], [7], [8], [9]. Despite their usefulness, PI formation requires specialized culture conditions and extensive cell manipulations.…”

Section: Discussionmentioning

confidence: 99%

“…The formation of 3-D β cell aggregates, or pseudoislets (PIs), is useful for the study of β cell biology. β-cells in PIs show improved function as measured by increased insulin production and improved glucose-stimulated insulin secretion (GSIS) [4], [5], [6], [7], [8], [9], [10]. These effects are mediated in part by the formation of a 3-D configuration which enhances β-cell – cell contact [5], [9], increases calcium signaling [11], and preserves extracellular matrix (ECM) proteins [12].…”

Section: Introductionmentioning

confidence: 99%

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In Vitro Formation of β Cell Pseudoislets Using Islet-Derived Endothelial Cells

et al. 2013

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β cell pseudoislets (PIs) are used for the in vitro study of β-cells in a three-dimensional (3-D) configuration. Current methods of PI induction require unique culture conditions and extensive mechanical manipulations. Here we report a novel co-culture system consisting of high passage β-cells and islet-derived endothelial cells (iECs) that results in a rapid and spontaneous formation of free-floating PIs. PI structures were formed as early as 72 h following co-culture setup and were preserved for more than 14 d. These PIs, composed solely of β-cells, were similar in size to that of native islets and showed an increased percentage of proinsulin-positive cells, increased insulin gene expression in response to glucose stimulation, and restored glucose-stimulated insulin secretion when compared to β-cells cultured as monolayers. Key extracellular matrix proteins that were absent in β-cells cultured alone were deposited by iECs on PIs and were found in and around the PIs. iEC-induced PIs are a readily available tool for examining β cell function in a native 3-D configuration and can be used for examining β-cell/iEC interactions in vitro.

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Section: Discussionsupporting

confidence: 48%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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In Vitro Formation of β Cell Pseudoislets Using Islet-Derived Endothelial Cells

et al. 2013

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“…Many attempts have been made to dissociate native islets and reaggregate them [ 20 – 26 ]. The most common method has employed spontaneous aggregation, where the dissociated islet cell suspension is cultured in ultra-low-binding plates [ 26 – 28 ].…”

Section: Introductionmentioning

confidence: 99%

Bioengineered human pseudoislets form efficiently from donated tissue, compare favourably with native islets in vitro and restore normoglycaemia in mice

Gamble

Pawlick

et al. 2018

Diabetologia

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Aims/hypothesisIslet transplantation is a treatment option that can help individuals with type 1 diabetes become insulin independent, but inefficient oxygen and nutrient delivery can hamper islet survival and engraftment due to the size of the islets and loss of the native microvasculature. We hypothesised that size-controlled pseudoislets engineered via centrifugal-forced-aggregation (CFA-PI) in a platform we previously developed would compare favourably with native islets, even after taking into account cell loss during the process.MethodsHuman islets were dissociated and reaggregated into uniform, size-controlled CFA-PI in our microwell system. Their performance was assessed in vitro and in vivo over a range of sizes, and compared with that of unmodified native islets, as well as islet cell clusters formed by a conventional spontaneous aggregation approach (in which dissociated islet cells are cultured on ultra-low-attachment plates). In vitro studies included assays for membrane integrity, apoptosis, glucose-stimulated insulin secretion assay and total DNA content. In vivo efficacy was determined by transplantation under the kidney capsule of streptozotocin-treated Rag1−/− mice, with non-fasting blood glucose monitoring three times per week and IPGTT at day 60 for glucose response. A recovery nephrectomy, removing the graft, was conducted to confirm efficacy after completing the IPGTT. Architecture and composition were analysed by histological assessment via insulin, glucagon, pancreatic polypeptide, somatostatin, CD31 and von Willebrand factor staining.ResultsCFA-PI exhibit markedly increased uniformity over native islets, as well as substantially improved glucose-stimulated insulin secretion (8.8-fold to 11.1-fold, even after taking cell loss into account) and hypoxia tolerance. In vivo, CFA-PI function similarly to (and potentially better than) native islets in reversing hyperglycaemia (55.6% for CFA-PI vs 20.0% for native islets at 500 islet equivalents [IEQ], and 77.8% for CFA-PI vs 55.6% for native islets at 1000 IEQ), and significantly better than spontaneously aggregated control cells (55.6% for CFA-PI vs 0% for spontaneous aggregation at 500 IEQ, and 77.8% CFA-PI vs 33.4% for spontaneous aggregation at 1000 IEQ; p < 0.05). Glucose clearance in the CFA-PI groups was improved over that in the native islet groups (CFA-PI 18.1 mmol/l vs native islets 29.7 mmol/l at 60 min; p < 0.05) to the point where they were comparable with the non-transplanted naive normoglycaemic control mice at a low IEQ of 500 IEQ (17.2 mmol/l at 60 min).Conclusions/interpretationThe ability to efficiently reformat dissociated islet cells into engineered pseudoislets with improved properties has high potential for both research and therapeutic applications.Electronic supplementary materialThe online version of this article (10.1007/s00125-018-4672-5) contains peer-reviewed but unedited supplementary material, which is available to authorised users.

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“…The mouse insulinoma cell line, MIN6 is a b-cell line known to possess the same vital characteristics as normal pancreatic b-cells and is a cost-effective replacement for primary islets in bcell research. 15,16 MIN6 b-cells are able to self organize into islet-like clusters called pseudoislets via reaggregation of single cells. These organized three-dimensional structures are important for the maintenance of normal insulin secretory responses as they closely mimic the native islet architecture, thus maintaining better cell-cell interactions.…”

Section: Introductionmentioning

confidence: 99%

Improving cellular function and immune protection via layer‐by‐layer nanocoating of pancreatic islet β‐cell spheroids cocultured with mesenchymal stem cells

Bhaiji

Zhi

Pickup

2012

J Biomedical Materials Res

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Islet transplantation as a therapy for type 1 diabetes is currently limited by lack of primary transplant material from human donors and post-transplantation loss of islets caused by adverse immune and nonimmune reactions. This study aimed to develop a novel strategy to create microenvironment for islets via integration of nanoencapsulation with cell cocultures, thereby enhancing their survival and function. The nanoencapsulation was achieved via layer-by-layer deposition of phosphorycholine-modified poly-L-lysine/heparin leading to the formation of nanometer-thick multilayer coating on islets. Spheroids formed by coculturing MIN6 β-cells with mesenchymal stem cells in suspension were used as the tool for testing encapsulation. Coculturing MSCs with MIN6 cells allowed the cell constructs to enhance structural and morphologic stability with improved insulin secretory function and render them less susceptible to inflammatory cytokine-induced apoptosis. Combining nanoencapsulation with coculture of MSCs/MIN6 resulted in higher glucose responsiveness, and lower antibody binding and apoptosis-inducing effects of cytokines. This strategy of nanoencapsulating islet cocultures appears promising to improve cellular delivery of insulin for treating type 1 diabetes.

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Pseudoislets as primary islet replacements for research

Cited by 16 publications

References 19 publications

In Vitro Formation of β Cell Pseudoislets Using Islet-Derived Endothelial Cells

In Vitro Formation of β Cell Pseudoislets Using Islet-Derived Endothelial Cells

Bioengineered human pseudoislets form efficiently from donated tissue, compare favourably with native islets in vitro and restore normoglycaemia in mice

Improving cellular function and immune protection via layer‐by‐layer nanocoating of pancreatic islet β‐cell spheroids cocultured with mesenchymal stem cells

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