Journal
JOVE-JOURNAL OF VISUALIZED EXPERIMENTS
Volume -, Issue 136, Pages -Publisher
JOURNAL OF VISUALIZED EXPERIMENTS
DOI: 10.3791/57327
Keywords
Immunology and Infection; Issue 136; Pancreatic islets; T2DM; reactive oxygen species; diabetogenic chemicals; ROS; xenobiotics
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Funding
- CREH center - NIEHS
- National Natural Science Foundation of China [31572626]
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Exposure to certain environmental chemicals in human and animals has been found to cause cellular damage of the pancreatic beta cells which will lead to the development of type 2 diabetes mellitus (T2DM). Although the mechanisms for the chemical-induced beta cell damage were unclear and likely to be complex, one recurring finding is that these chemicals induce oxidative stress leading to the generation of excessive reactive oxygen species (ROS) which induce damage to the beta cell. To identify potential diabetogenic environmental chemicals, we isolated pancreatic islet cells from C57BL/6 mice and cultured islet cells in 96-well cell culture plates; then, the islet cells were dosed with chemicals and the ROS generation was detected by 2',7'-dichlorofluorescein (DCFH-DA) fluorescent dye. Using this method, we found that bisphenol A (BPA), Benzo[a]pyrene (BaP), and polychlorinated biphenyls (PCBs), could induce high levels of ROS, suggesting that they may potentially induce damage in islet cells. This method should be useful for screening diabetogenic xenobiotics. In addition, the cultured islet cells may also be adapted for in vitro analysis of chemical-induced toxicity in pancreatic cells.
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