A characterization of the ZFL cell line and primary hepatocytes as in vitro liver cell models for the zebrafish (Danio rerio)
A characterization of the ZFL cell line and primary hepatocytes as in vitro liver cell models for the zebrafish (Danio rerio)
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DOI：
10.1016/j.aquatox.2013.11.023
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发表时间：
2014-02-01
影响因子：
4.5
通讯作者：
Goksoyr, Anders
中科院分区：
文献类型：
--
作者：
Eide, Marta;Rusten, Marte;Goksoyr, Anders
斑马鱼（Danio redo）是生物医学研究中广泛使用的模式生物。从斑马鱼肝脏建立的ZFL细胞系和从斑马鱼新鲜分离的原代肝细胞已用于几项毒理学研究。然而，没有以前的报告进行了比较，并在基因表达水平的特点，这两个系统。本研究的目的是评估ZFL细胞系相比，原代肝细胞作为体外模型研究环境污染物对斑马鱼肝脏的影响。采用实时荧光定量PCR技术，比较了斑马鱼ZFL细胞系、原代肝细胞和全肝中参与毒性反应的关键基因的基础水平和转录诱导潜力。研究表明，与斑马鱼肝脏相比，ZFL细胞中大多数选定基因的mRNA水平较低。在测试剂量下，与斑马鱼原代肝细胞相比，ZFL细胞暴露于配体后诱导的基因转录要低得多。重要的是，雌激素受体和卵黄蛋白原基因表现出较低的基础转录，在ZFL细胞系中没有诱导反应。总之，原代肝细胞似乎非常适合研究环境污染物，包括外源性雌激素，但可能会显示大的性别依赖性差异的基因转录。ZFL细胞系在涉及芳烃受体途径的毒理学研究中显示出潜力。然而，一般来说，基因转录诱导的可能性很低，特别是在研究雌激素反应时。(C)2013爱思唯尔有限公司版权所有。
The zebrafish (Danio redo) is a widely used model species in biomedical research. The ZFL cell line, established from zebrafish liver, and freshly isolated primary hepatocytes from zebrafish have been used in several toxicological studies. However, no previous report has compared and characterized these two systems at the level of gene expression. The aim of this study was to evaluate the ZFL cell line in comparison to primary hepatocytes as in vitro models for studying effects of environmental contaminants in zebrafish liver. Using quantitative real-time PCR, the basal level and transcriptional induction potential of key genes involved in toxic responses in the ZFL cell line, primary hepatocytes and whole liver from zebrafish were compared. The study showed that the ZFL cells have lower levels of mRNA of most selected genes compared to zebrafish liver. The induced gene transcription following exposure to ligand was much lower in ZFL cells compared to zebrafish primary hepatocytes at the doses tested. Importantly, oestrogen receptor and vitellogenin genes showed low basal transcription and no induction response in the ZFL cell line. In conclusion, it appears that primary hepatocytes are well suited for studying environmental contaminants including xenoestrogens, but may show large sex-dependent differences in gene transcription. The ZFL cell line shows potential in toxicological studies involving the aryl hydrocarbon receptor pathway. However, low potential for transcriptional induction of genes in general should be expected, especially notable when studying estrogenic responses. (C) 2013 Elsevier B.V. All rights reserved.