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Assessment of 16 chemicals on proliferation and apoptosis in human neuroprogenitor cells using high-content image analysis (HCA).
Citation:
Culbreth, M. E., J. HARRILL, T. M. Freudenrich, W. R. MUNDY, AND TIM SHAFER. Assessment of 16 chemicals on proliferation and apoptosis in human neuroprogenitor cells using high-content image analysis (HCA). Presented at Society of Toxicology (SOT) Annual Meeting, Washington, NC, March 06 - 10, 2011.
Impact/Purpose:
The need for efficient methods of screening chemicals for the potential to cause developmental neurotoxicity is paramount
Description:
The need for efficient methods of screening chemicals for the potential to cause developmental neurotoxicity is paramount. We previously described optimization of an HCA assay for proliferation and apoptosis in ReNcell CX cells (ReN), identifying appropriate controls. Utility of the assay was examined using 16 chemicals: Cd, chlorpyrifos oxon (CPO), dexamethasone, dieldrin, ketamine, Pb, maneb, methylmercury (MeHg), nicotine, trans-retinoic acid (tRA), and trimethyltin (TMT) reported to be developmentally neurotoxic and to affect proliferation and apoptosis; Dimethyl phthalate, glyphosate, omeprazole, saccharin, and sorbitol not reported to be developmentally neurotoxic or affect proliferation and apoptosis below 30uM. Cells in 96-well plates were exposed (0.001-1 OOuM, N=3) for 24h, and proliferation and apoptosis assessed using HCA based on BrdU incorporation and activation of p53 and caspase 3, respectively. Cell viability was measured using Cell Titer-GLO(TM). Noneof the 16 chemicals significantly actlvated p53 or caspase 3 compared to actinomycin d, which increased these by ~1000%. Thus, whether these chemicals cause apoptosis in ReN cells through other pathways needs to be assessed. Compared to control, Cd, Pb, MeHg, TMT and tRA decreased BrdU by 50-80%; CPO, dieldrin, maneb, and nicotine decreased BrdU by 30-50%; dexamethasone, ketamine, dimethyl phthalate, glyphosate, omeprazole, saccharin, and sorbitol decreased BrdU by 10-30%. At the highest concentration tested, Cd, Pb, MeHg, TMT, and tRA decreased viability by >50%, while dieldrin decreased by it by ~30%; all other chemicals had no effect on viability. Effects on proliferation were always observed at lower concentrations than effects on viability. The results show that BrdU incorporation is sensitive to chemical effects on neuroprogenitor cell proliferation.This abstract does not reflect Agency Policy.