<i>In vitro</i>
            cultivation of human islets from expanded ductal tissue

Bonner-Weir, Susan; Taneja, Monica; Weir, Gordon C.; Tatarkiewicz, Krystyna; Song, Ki‐Ho; Sharma, Arun; O’Neil, John J.

doi:10.1073/pnas.97.14.7999

Cited by 924 publications

(660 citation statements)

References 27 publications

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“…Activation of Pdx1 in duct cells of the pancreas has also been observed following partial pancreatectomy in mice [85] and in insulin resistant Zucker fatty (ZF) rats [83]. Importantly in this regard, Pdx1 expression is often observed in proliferating duct cells [77,94,95], and such cells have been suggested to serve as substrate for β cell neogenesis, particularly in cases of adaptive hyperplasia or pancreas regeneration [92,[96][97][98][99]. A recent study suggests that Pdx1, in complex with TALE transcription factors, negatively regulates the duct-specific keratin 19 gene in mature duct cells [100].…”

Section: Role Of Pdx1 In Adaptive β Cell Hyperplasia and β Cell Regenmentioning

confidence: 99%

A feat of metabolic proportions: Pdx1 orchestrates islet development and function in the maintenance of glucose homeostasis

Babu

Deering

Mirmira

2007

Molecular Genetics and Metabolism

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Emerging evidence over the past decade indicates a central role for transcription factors in the embryonic development of pancreatic islets and the consequent maintenance of normal glucose homeostasis. Pancreatic and duodenal homeobox 1 (Pdx1) is the best studied and perhaps most important of these factors. Whereas deletion or inactivating mutations of the Pdx1 gene causes whole pancreas agenesis in both mice and humans, even haploinsufficiency of the gene or alterations in its expression in mature islet cells causes substantial impairments in glucose tolerance and the development of a late-onset form of diabetes known as maturity onset diabetes of the young. The study of Pdx1 has revealed crucial phenotypic interrelationships of the varied cell types within the pancreas, particularly as these impinge upon cellular differentiation in the embryo and neogenesis and regeneration in the adult. In this review, we describe the actions of Pdx1 in the developing and mature pancreas and attempt to unify these actions with its known roles in modulating transcriptional complex formation and chromatin structure at the molecular genetic level.

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Section: Role Of Pdx1 In Adaptive β Cell Hyperplasia and β Cell Regenmentioning

confidence: 99%

A feat of metabolic proportions: Pdx1 orchestrates islet development and function in the maintenance of glucose homeostasis

Babu

Deering

Mirmira

2007

Molecular Genetics and Metabolism

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“…Primary antibody concentrations were as follows: 1/1,000 guinea pig anti-porcine insulin (Dako, Glostrup, Denmark), 1/50 mouse anti-human CK19 (Dako), 1/200 mouse anti-human CK7 (Dako). Antibodies to CK7 and 19 were used as markers for pancreatic ductal epithelial-type cells as previously described [33]. All biotinylated secondary antibodies were obtained from Vector Laboratories (Burlingame, CA, USA) and used at a concentration of 1/200.…”

Section: Cell Characterisationmentioning

confidence: 99%

The impact of the mTOR inhibitor sirolimus on the proliferation and function of pancreatic islets and ductal cells

et al. 2006

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Aims/hypothesis The Edmonton Protocol for islet transplantation has provided hope for type 1 diabetic patients. However, this protocol requires lifelong immunosuppression, specifically sirolimus, a cellular antiproliferate. The effect of sirolimus on human pancreatic ductal cells (HDCs) is not known. This may be important since HDCs are believed to be islet precursors. Since neonatal porcine islets (NPIs), which contain many ductal precursor cells, could be a potential clinical source of islets, we also tested the effects of sirolimus on this tissue. Methods HDCs (n=4), NPIs (n=9) and human islets (n=5) were cultured with and without sirolimus (20 ng/ml) for 6 days. Results HDCs and NPIs cultured with sirolimus showed a 50 and 28% decrease, respectively, in cell number relative to control (p<0.05). Control cultures expanded 1.65-and 2.44-fold relative to time 0. Decreases in cell number of sirolimus-treated HDCs were not due to apoptosis as measured by TUNEL staining. No functional effects on human islets or NPIs were observed following static incubation with high glucose. Treatment of syngeneically transplanted and naïve BALC/c mice with sirolimus resulted in altered OGTT profiles with prolonged elevation of hyperglycaemia and weight gain. There was no difference in graft and organ insulin content between treatment groups. Conclusions/interpretation Our results indicate that sirolimus decreases ductal cell numbers in culture and alters glucose-stimulated insulin secretion in vivo. The administration of sirolimus to islet transplant recipients is likely to impair graft function as a result of decreasing ductal neogenesis and induction of insulin resistance.

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“…Optical density values (OD; 450 nm) were corrected for background levels. Nuclear extracts were obtained after indicated time intervals for co-culture of 2.5×10 5 Capan-2 cells alone (black histogram), 2.5×10 5 Capan-2 cells co-cultured with 2.5×10 5 CD40L-transfected fibroblasts (3T6 CD40L ; white histogram) or 2.5×10 5 Capan-2 cells co-cultured with 2.5×10 5 untransfected fibroblasts (3T6 CTRL ; hatched histogram). One representative experiment out of three is shown.…”

Section: Cd40 Engagement Induces Tnf-α and Il-1β Mrna Expressionmentioning

confidence: 99%

“…There is suggestive evidence that pancreatic ducts are potential sites for beta cell neogenesis [1,2,3,4,5,6,7]. Duct cells were also shown to produce nitric oxide [4] and to express MHC class II [3], leading to proposals that they might contribute to beta cell damage in Type 1 diabetes mellitus [8].…”

Section: Introductionmentioning

confidence: 99%

CD40 expression on human pancreatic duct cells: role in nuclear factor-kappa B activation and production of pro-inflammatory cytokines

Vosters

Beuneu

Nagy³

et al. 2004

Diabetologia

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Aims/hypothesis. Human pancreatic duct cells are closely associated with islet beta cells, and contaminate islet suspensions transplanted in Type 1 diabetes mellitus patients. Activated duct cells produce cytotoxic mediators and possibly contribute to the pathogenesis of Type 1 diabetes mellitus or islet graft rejection. As CD40 transduces activation signals involved in inflammatory and immune disorders, we investigated CD40 expression on duct cells and their response to CD40 engagement. Methods. CD40 expression on human pancreatic duct cells was analysed by flow cytometry and immunohistochemical analyses. To assess the function of CD40 expression on duct cells, activation of the transcription factor nuclear factor-kappa B was determined using electrophoretic mobility shift assay and ELISA. Cytokine mRNA levels were quantified by real-time RT-PCR, and protein levels by Luminex technology. Results. Isolated human pancreatic duct cells and Capan-2 cell lines were found to express constitutively CD40. The expression of CD40 on duct cells was confirmed in vivo on human normal and pathological pancreatic specimens. CD40 ligation on Capan-2 cells induced rapid nuclear factor-kappa B activation, and supershift assays demonstrated that p50/p65 heterodimers and p50/p50 homodimers were present in the activated complexes in the nucleus. This activation was accompanied by tumour necrosis factor-α and interleukin-1β mRNA accumulation. Tumour necrosis factor-α protein secretion was confirmed in CD40-activated Capan-2 cells and in isolated human pancreatic duct cells. Conclusions/interpretation. Interaction between activated T lymphocytes expressing CD40 ligand and duct cells expressing CD40 may contribute to the immune responses involved in Type 1 diabetes mellitus and islet graft rejection. Interfering with CD40-mediated duct cell activation could alleviate beta cell damage of immune origin.

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In vitro cultivation of human islets from expanded ductal tissue

Cited by 924 publications

References 27 publications

A feat of metabolic proportions: Pdx1 orchestrates islet development and function in the maintenance of glucose homeostasis

A feat of metabolic proportions: Pdx1 orchestrates islet development and function in the maintenance of glucose homeostasis

The impact of the mTOR inhibitor sirolimus on the proliferation and function of pancreatic islets and ductal cells

CD40 expression on human pancreatic duct cells: role in nuclear factor-kappa B activation and production of pro-inflammatory cytokines

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