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A Human Islet Cell Culture System for High-Throughput Screening.
|Publication Type||Journal Article|
|Authors||Walpita, D., Hasaka T., Spoonamore J., Vetere A., Takane KK, Fomina-Yadlin D., Fiaschi-Taesch N., Shamji A., Clemons PA, Stewart AF, Schreiber SL, and Wagner BK|
|Abstract||A small-molecule inducer of beta-cell proliferation in human islets represents a potential regeneration strategy for treating type 1 diabetes. However, the lack of suitable human beta cell lines makes such a discovery a challenge. Here, we adapted an islet cell culture system to high-throughput screening to identify such small molecules. We prepared microtiter plates containing extracellular matrix from a human bladder carcinoma cell line. Dissociated human islets were seeded onto these plates, cultured for up to 7 days, and assessed for proliferation by simultaneous Ki67 and C-peptide immunofluorescence. Importantly, this environment preserved beta-cell physiological function, as measured by glucose-stimulated insulin secretion. Adenoviral overexpression of cdk-6 and cyclin D(1), known inducers of human beta cell proliferation, was used as a positive control in our assay. This induction was inhibited by cotreatment with rapamycin, an immunosuppressant often used in islet transplantation. We then performed a pilot screen of 1280 compounds, observing some phenotypic effects on cells. This high-throughput human islet cell culture method can be used to assess various aspects of beta-cell biology on a relatively large number of compounds.|
|Year of Publication||2011|
|Journal||Journal of biomolecular screening|
|Date Published (YYYY/MM/DD)||2011/12/07|