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DOI | 10.1016/j.tiv.2016.08.006 |
Development of an in vitro high content imaging assay for quantitative assessment of CAR-dependent mouse, rat, and human primary hepatocyte proliferation | |
Soldatow, Valerie1; Peffer, Richard C.2; Trask, O. Joseph1; Cowie, David E.2; Andersen, Melvin E.1,3; LeCluyse, Edward1; Deisenroth, Chad1,3,4 | |
发表日期 | 2016-10-01 |
ISSN | 0887-2333 |
卷号 | 36页码:224-237 |
英文摘要 | Rodent liver tumors promoted by constitutive androstane receptor (CAR) activation are known to be mediated by key events that include CAR-dependent gene expression and hepatocellular proliferation. Here, an in vitro high content imaging based assay was developed for quantitative assessment of nascent DNA synthesis in primary hepatocyte cultures from mouse, rat, and human species. Detection of DNA synthesis was performed using direct DNA labeling with the nucleoside analog 5-ethynyl-2'-deoxyuridine (EdU). The assay was multiplexed to enable direct quantitation of DNA synthesis, cytotoxicity, and cell count endpoints. An optimized defined medium cocktail was developed to sensitize hepatocytes to cell cycle progression. The baseline EdU response to defined medium was greatest for mouse, followed by rat, and then human. Hepatocytes from all three species demonstrated CAR, activation in response to the CAR agonists TCPOBOP, CITCO, and phenobarbital based on increased gene expression for Cyp2b isoforms. When evaluated for a proliferation phenotype, TCPOBOP and CITCO exhibited significant dose-dependent increases in frequency of EdU labeling in mouse and rat hepatocytes that was not observed in hepatocytes from three human donors. The observed species differences are consistent with CAR activators inducing a proliferative response in rodents, a key event in the liver tumor mode of action that is not observed in humans. (C) 2016 Elsevier Ltd. All rights reserved. |
英文关键词 | Primary hepatocytes;Cell proliferation;High content imaging;Car |
语种 | 英语 |
WOS记录号 | WOS:000383826000026 |
来源期刊 | TOXICOLOGY IN VITRO |
来源机构 | 美国环保署 |
文献类型 | 期刊论文 |
条目标识符 | http://gcip.llas.ac.cn/handle/2XKMVOVA/59645 |
作者单位 | 1.Hamner Inst Hlth Sci, Inst Chem Safety Sci, 6 Davis Dr,POB 12137, Res Triangle Pk, NC 27709 USA; 2.Syngenta Crop Protect LLC, POB 18300,410 Swing Rd, Greensboro, NC 27419 USA; 3.ScitoVation LLC, 6 Davis Dr,POB 110566, Res Triangle Pk, NC 27709 USA; 4.US EPA, Natl Ctr Computat Toxicol, Res Triangle Pk, NC 27709 USA |
推荐引用方式 GB/T 7714 | Soldatow, Valerie,Peffer, Richard C.,Trask, O. Joseph,et al. Development of an in vitro high content imaging assay for quantitative assessment of CAR-dependent mouse, rat, and human primary hepatocyte proliferation[J]. 美国环保署,2016,36:224-237. |
APA | Soldatow, Valerie.,Peffer, Richard C..,Trask, O. Joseph.,Cowie, David E..,Andersen, Melvin E..,...&Deisenroth, Chad.(2016).Development of an in vitro high content imaging assay for quantitative assessment of CAR-dependent mouse, rat, and human primary hepatocyte proliferation.TOXICOLOGY IN VITRO,36,224-237. |
MLA | Soldatow, Valerie,et al."Development of an in vitro high content imaging assay for quantitative assessment of CAR-dependent mouse, rat, and human primary hepatocyte proliferation".TOXICOLOGY IN VITRO 36(2016):224-237. |
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