Characterization of Endocrine Progenitor Cells and Critical Factors for Their Differentiation in Human Adult Pancreatic Cell Culture

Gao, Ru; Ustinov, Jarkko; Pulkkinen, Mari‐Anne; Lundin, Karolina; Korsgren, Olle; Otonkoski, Timo

doi:10.2337/diabetes.52.8.2007

Cited by 221 publications

(161 citation statements)

References 52 publications

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“…Following isolation, the remaining exocrine fractions were examined and those that were duct-rich but islet-depleted [24,25] were collected and washed in sterile Hanks' balanced salt solution (Sigma-Aldrich) prior to being plated into T75 flasks (Fisher Scientific, Loughborough, UK). The exocrine cultures were maintained undisturbed for 48 h at 37°C, in an atmosphere of 95% O 2 and 5% CO 2 , in Dulbecco's modified Eagle's medium supplemented with 10% FBS (Sigma-Aldrich), 100 U/ml penicillin, 100 µg/ml streptomycin, 10 µg/ml amphotericin B. Adherent tissue remaining at the time of the first medium change was allowed to expand further to form an 80% confluent monolayer over the following 48-72 h.…”

Section: Preparation Of Dec Culturesmentioning

confidence: 99%

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Sustained insulin secretory response in human islets co-cultured with pancreatic duct-derived epithelial cells within a rotational cell culture system

et al. 2009

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Aims/hypothesis Loss of the trophic support provided by surrounding non-endocrine pancreatic cell populations underlies the decline in beta cell mass and insulin secretory function observed in human islets following isolation and culture. This study sought to determine whether restoration of regulatory influences mediated by ductal epithelial cells promotes sustained beta cell function in vitro. Methods Human islets were isolated according to existing protocols. Ductal epithelial cells were harvested from the exocrine tissue remaining after islet isolation, expanded in monolayer culture and characterised using fluorescence immunocytochemistry. The two cell types were co-cultured under conventional static culture conditions or within a rotational cell culture system. The effect of co-culture on islet structural integrity, beta cell mass and insulin secretory capacity was observed for 10 days following isolation. Results Human islets maintained under conventional culture conditions exhibited a characteristic loss in structural integrity and functional viability as indicated by a diminution of glucose responsiveness. By contrast, co-culture of islets with ductal epithelial cells led to preserved islet morphology and sustained beta cell function, most evident in co-cultures held within the rotational cell culture system, which showed a significantly (p<0.05) greater insulin secretory response to elevated glucose compared with control islets. Similarly, insulin/protein ratio data suggested that the presence of ductal epithelial cells is beneficial for the maintenance of beta cell mass. Conclusions/interpretation The data indicate a supportive role for ductal epithelial cells in islet viability. Further characterisation of the regulatory influences may lead to novel strategies to improve long-term beta cell function both in vitro and following islet transplantation.

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Section: Preparation Of Dec Culturesmentioning

confidence: 99%

“…The islet-depleted exocrine fraction was washed and plated for culture at high density in T75 flasks as previously described [21,24]. The vast majority (>80%) of the plated material remained non-adherent at 48 h and was discarded in the first medium change.…”

Section: Morphological and Immunocytochemical Assessment Of Islet Andmentioning

confidence: 99%

Sustained insulin secretory response in human islets co-cultured with pancreatic duct-derived epithelial cells within a rotational cell culture system

et al. 2009

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“…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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“…An in vitro-generated islet buds released insulin in response to glucose nearly as efficiently as native islets. Although, when transplanted under the kidney capsule of nude mice, only one of five grafts demonstrated further growth with foci of both endocrine and exocrine differentiation (Gao et al 2003). Liver stem/progenitor cells are self-renewing capability and multilineage differentiation potential (Suzuki et al 2002).…”

Section: Differentiation Of Progenitor Cellsmentioning

confidence: 99%

“…Although stem cells were isolated from pancreatic ducts, islets and exocrine tissue the b-cell progenitors have not been identified (Baeyens et al 2005;Carlotti et al 2010;Gao et al 2003;Soria et al 2005). Interestingly, b cells mass increases in vivo significantly after injury and during metabolic demand for example pregnancy or obesity.…”

Section: Introductionmentioning

confidence: 99%

Differentiation of Stem Cells into Insulin-Producing Cells: Current Status and Challenges

Pokrywczyńska

Krzyzanowska

Jundziłł

et al. 2013

Arch. Immunol. Ther. Exp.

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Diabetes mellitus is one of the most serious public health challenges of the twenty-first century. Allogenic islet transplantation is an efficient therapy for type 1 diabetes. However, immune rejection, side effects of immunosuppressive treatment as well as lack of sufficient donor organs limits its potential. In recent years, several promising approaches for generation of new pancreatic b cells have been developed. This review provides an overview of current status of pancreatic and extra-pancreatic stem cells differentiation into insulin-producing cells and the possible application of these cells for diabetes treatment. The PubMed database was searched for English language articles published between 2001 and 2012, using the keyword combinations: diabetes mellitus, differentiation, insulin-producing cells, stem cells.

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Characterization of Endocrine Progenitor Cells and Critical Factors for Their Differentiation in Human Adult Pancreatic Cell Culture

Cited by 221 publications

References 52 publications

Sustained insulin secretory response in human islets co-cultured with pancreatic duct-derived epithelial cells within a rotational cell culture system

Sustained insulin secretory response in human islets co-cultured with pancreatic duct-derived epithelial cells within a rotational cell culture system

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

Differentiation of Stem Cells into Insulin-Producing Cells: Current Status and Challenges

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