Transcriptional profiling of stress-response in cultured porcine islets
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ABSTRACT: An increasing demand for cell-based diabetes therapy could be met through xenotransplantation of adult porcine islets. Use of islet xenotranplantation on a large scale would require rigorous safety and quality control measures to maximize transplant success. Development of molecular tools to monitor porcine islet cellular responses to ischemic, osmotic, mechanical and oxidative stresses during islet cell processing and post-isolation culturing would aid the rational design of cytoprotective strategies aimed at improving transplant outcomes. In addition, gene expression signatures informative for islet quality could serve as an adjunct to physiological testing to establish the suitability of islet products for transplantation. Nine adult Landrace sows (2-3.5 y old, 249±27 kg) were sacrificed, the pancreases were dissected, and islet cells isolated as previously described [8]. Islet preparation purity was assessed by light microscopy after staining with diphenylthiocarbazone and ranged between 90-95% for the preparations used. Islet yield, based on islet equivalents (IEQ, number of islets standardized to 150 µm diameter), was estimated at 1930±520 per gram of pancreas tissue.Profiles of islet cells cultured under standard conditions were compared to islet cells cultured under stress conditions with elevated glucose (16.7 mM) or addition of inflammatory cytokines (IL-1ï¢, TNF-ï¡, and IFN-ï§), or both, for 48 hours.
ORGANISM(S): Sus scrofa
SUBMITTER: M.P. Murtaugh
PROVIDER: E-GEOD-4744 | biostudies-arrayexpress |
REPOSITORIES: biostudies-arrayexpress
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