A New Culture Method for Human Pancreatic Islets Using a Biopore Membrane Insert

Hober, C; Py, Benhamou; Pc, Watt; Watanabe, Yasumasa; Nomura, Yuji; Stein, Elizabeth; Fc, Brunicardi; Mullen, Y

doi:10.1097/00006676-199703000-00014

Cited by 14 publications

(7 citation statements)

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“…In the current study, the purity of islet culture was increased (Ͼ80%) by reducing contaminating exocrine tissue and ductal fragments by islet handpicking, and to avoid fibroblast contamination, pancreatic islets were cultured with noncoated transparent membrane inserts. Since the membrane does not allow cell attachment or fibroblast growth, islets cultured in the insert maintained their original structure and remained in a free-floating form after long-term culture (22). Virus infection was carried out with intact islets rather than dissociated cells, because tissues that are resistant to virus infections in vivo can artificially be made susceptible by dispersing the cells into culture (40).…”

Section: Discussionmentioning

confidence: 99%

“…Islet cultures in noncoated membrane inserts have been shown to prevent fibroblast growth and to maintain normal islet function (22). In our islet culture system, we used Nunc tissue culture 8-well strip inserts (A/S Nunc, Roskilde, Denmark) and 96-well plate inserts to study IFN-␣ production and Millicell-CM (12 mm in diameter; Millipore, Bedford, Mass.)…”

Section: Methodsmentioning

confidence: 99%

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Persistent Infection of Human Pancreatic Islets by Coxsackievirus B Is Associated with Alpha Interferon Synthesis in β Cells

Chehadeh¹,

Kerr–Conte²,

Pattou³

et al. 2000

J Virol

176

147

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Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Persistent Infection of Human Pancreatic Islets by Coxsackievirus B Is Associated with Alpha Interferon Synthesis in β Cells

Chehadeh¹,

Kerr–Conte²,

Pattou³

et al. 2000

J Virol

176

147

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“…The labelled cells (2·5 × 10 5 /ml) were incubated for 48 hr with irradiated A20‐HL cells (5 × 10 5 /ml) using a 6‐ml round‐bottom tube (Falcon 2054; Becton Dickinson and Company, Franklin Lake, NJ). Alternatively, BCECF‐labelled cells (2·5 × 10 5 /ml) were co‐cultured for 48 hr with irradiated A20‐HL cells (5 × 10 5 cells/ml) isolated in Millicell chamber containing a 0·45‐µm nitrocellulose filter (Millipore Co., Bedford, MA) on a 24‐well culture plate 26 . They were analysed for CD80 expression on a FACSCalibur.…”

Section: Methodsmentioning

confidence: 99%

“…45-mm nitrocellulose filter (Millipore Co., Bedford, MA) on a 24-well culture plate. 26 They were analysed for CD80 expression on a FACSCalibur.…”

Section: Co-culture Assaymentioning

confidence: 99%

Irradiation up‐regulates CD80 expression through induction of tumour necrosis factor‐α and CD40 ligand expression on B lymphoma cells

Ishikawa

Nakano²,

Seo³

et al. 2002

Immunology

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SUMMARYPreviously, we reported that 100 Gy X-ray irradiation followed by 24 hr incubation up-regulates CD80 expression in murine B lymphoma cells, A20-2J. In the present study, we analysed the underlying mechanisms of such up-regulation using A20-HL cells derived from A20-2J cells. Irradiation of A20-HL cells with 100 Gy enhanced CD80 expression. Incubation of untreated A20-HL cells with those 100 Gy irradiated induced up-regulation of CD80 expression. Irradiation of A20-HL cells also up-regulated the expression of tumour necrosis factor-a (TNF-a) and CD40 ligand (CD40L), and the amount of immunoprecipitable TNF-a and CD40L in cell lysates. The addition of anti-TNF-a or anti-CD40L monoclonal antibody (mAb) to the incubation of irradiated A20-HL cells partially inhibited up-regulation of CD80 expression, and the addition of both antibodies together almost completely inhibited the up-regulation, suggesting that irradiation up-regulated the CD80 expression through the induction of TNF-a and CD40L expression. Irradiation also increased the accumulation of CD80, TNF-a and CD40L mRNA. n-tosyl-L-phenylalanine chloromethyl ketone (TPCK), a nuclear factor (NF)-kB inhibitor, markedly decreased irradiation-induced accumulation of CD80 mRNA and CD80 expression. FK506, a calcineurin inhibitor, and nifedipine, a calcium channel inhibitor, inhibited not only the expression of TNF-a and CD40L, but also the up-regulation of CD80 on irradiated A20-HL cells. These results strongly suggested that irradiation induced TNF-a and CD40L expression, which then up-regulated CD80 mRNA and CD80 expression through activation of NF-kB transcription factor in A20-HL cells.

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“…Isolation of human islet cells has so far proved to be difficult and long-term survival under in vitro culture conditions has been limited 25–31. As observed with ductal cells, cultured islet cells undergo senescence in in vitro culture.…”

Section: In Vitro Culture Of Non-neoplastic Pancreatic Cellsmentioning

confidence: 99%

In vitromodels of pancreatic cancer for translational oncology research

Feldmann

Rauenzahn

Maitra

2009

Expert Opinion on Drug Discovery

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Background-Pancreatic cancer is a disease of near uniform fatality and the overwhelming majority of patients succumb to their advanced malignancy within a few months of diagnosis. Despite considerable advances in our understanding of molecular mechanisms underlying pancreatic carcinogenesis, this knowledge has not yet been fully translated into clinically available treatment strategies that yield significant improvements in disease free or overall survival.Objective-Cell line-based in vitro model systems provide powerful tools to identify potential molecular targets for therapeutic intervention as well as for initial pre-clinical evaluation of novel drug candidates. Here we provide a brief overview of recent literature on cell line-based model systems of pancreatic cancer and their application in the search for novel therapeutics against this vicious disease.Conclusion-While in vitro models of pancreatic cancer are of tremendous value for genetic studies and initial functional screenings in drug discovery, they carry several imanent drawbacks and are often poor in predicting therapeutic response in humans. Therefore, in most instances they are successfully exploited to generate hypothesis and identify molecular targets for novel therapeutics, which are subsequently subject to further in-depth characterization using more advanced in vivo model systems and clinical trials.

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A New Culture Method for Human Pancreatic Islets Using a Biopore Membrane Insert

Cited by 14 publications

References 0 publications

Persistent Infection of Human Pancreatic Islets by Coxsackievirus B Is Associated with Alpha Interferon Synthesis in β Cells

Persistent Infection of Human Pancreatic Islets by Coxsackievirus B Is Associated with Alpha Interferon Synthesis in β Cells

Irradiation up‐regulates CD80 expression through induction of tumour necrosis factor‐α and CD40 ligand expression on B lymphoma cells

In vitromodels of pancreatic cancer for translational oncology research

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