跳转至内容
Merck
  • Evaluation of the sensitivity, responsiveness and reproducibility of primary rainbow trout hepatocyte vitellogenin expression as a screening assay for estrogen mimics.

Evaluation of the sensitivity, responsiveness and reproducibility of primary rainbow trout hepatocyte vitellogenin expression as a screening assay for estrogen mimics.

Aquatic toxicology (Amsterdam, Netherlands) (2015-01-07)
Maria T Hultman, Jan Thomas Rundberget, Knut Erik Tollefsen
摘要

Vitellogenin (Vtg) induction in primary fish hepatocytes has been proposed as an in vitro screening assay for ER agonists and antagonists, but has not yet been used extensively as a high-throughput screening tool due to poor reproducibility, sensitivity and overall feasibility. The present work has evaluated the role of seasonal variation, normalization, optimal culture and assay conditions on the sensitivity, responsiveness and reproducibility of in vitro vtg gene mRNA and protein expression in rainbow trout (Oncorhynchus mykiss) primary hepatocytes using the xenoestrogen 17α-ethynylestradiol (EE2) as a test chemical. The results show that primary hepatocytes display a relatively high individual and seasonal variation in both Vtg mRNA and protein induction potential, although less variance was observed in assay sensitivity. Data normalization of assay response to maximum (3 nM EE2) and minimum (DMSO) Vtg production dramatically reduced this variance and led to improved assay reproducibility. A time-dependent response was observed both for mRNA and protein expression, reaching maximum Vtg induction after 96 h of exposure, although reproducible concentration response curves for both Vtg mRNA and protein could be obtained already after 48 h. A need for chemical re-exposure of the hepatocytes was identified to be important for sustaining exposure concentrations in extended studies (>48 h), whereas different plate formats (96, 24 or 6 wells) did not affect the bioassay performance. In conclusion, standardization of hepatocyte bioassay and test conditions as well as data-normalization procedures are proposed to be instrumental for more consistent and comparable results in future use of this in vitro assay.

材料
货号
品牌
产品描述

Sigma-Aldrich
甲醇, suitable for HPLC, ≥99.9%
价格与库存信息目前不能提供
Sigma-Aldrich
丙酮, ACS reagent, ≥99.5%
价格与库存信息目前不能提供
Sigma-Aldrich
甲醇, ACS reagent, ≥99.8%
价格与库存信息目前不能提供
Sigma-Aldrich
丙酮, suitable for HPLC, ≥99.9%
价格与库存信息目前不能提供
Sigma-Aldrich
甲醇, suitable for HPLC, gradient grade, ≥99.9%
价格与库存信息目前不能提供
Sigma-Aldrich
丙酮, HPLC Plus, for HPLC, GC, and residue analysis, ≥99.9%
价格与库存信息目前不能提供
Sigma-Aldrich
甲苯, ACS reagent, ≥99.5%
价格与库存信息目前不能提供
Sigma-Aldrich
碳酸氢钠, ACS reagent, ≥99.7%
价格与库存信息目前不能提供
Sigma-Aldrich
甲醇, HPLC Plus, ≥99.9%
价格与库存信息目前不能提供
Sigma-Aldrich
无水碳酸钠, powder, ≥99.5%, ACS reagent
价格与库存信息目前不能提供
Sigma-Aldrich
甲苯, suitable for HPLC, 99.9%
价格与库存信息目前不能提供
Sigma-Aldrich
甲苯, HPLC Plus, for HPLC, GC, and residue analysis, ≥99.9%
价格与库存信息目前不能提供
Sigma-Aldrich
碳酸氢钠, powder, BioReagent, for molecular biology, suitable for cell culture, suitable for insect cell culture
价格与库存信息目前不能提供
Sigma-Aldrich
丙酮, Laboratory Reagent, ≥99.5%
价格与库存信息目前不能提供
Sigma-Aldrich
乙酸铵, ACS reagent, ≥97%
价格与库存信息目前不能提供
Sigma-Aldrich
无水碳酸钠, ACS reagent, anhydrous, ≥99.5%, powder or granules
价格与库存信息目前不能提供
Sigma-Aldrich
碳酸氢钠, ReagentPlus®, ≥99.5%, powder
价格与库存信息目前不能提供
Sigma-Aldrich
乙酸铵, ≥99.99% trace metals basis
价格与库存信息目前不能提供
Sigma-Aldrich
硫酸铜, ReagentPlus®, ≥99%
价格与库存信息目前不能提供
Sigma-Aldrich
甲醇, puriss. p.a., ACS reagent, reag. ISO, reag. Ph. Eur., ≥99.8% (GC)
价格与库存信息目前不能提供
Sigma-Aldrich
甲苯, puriss. p.a., ACS reagent, reag. ISO, reag. Ph. Eur., ≥99.7% (GC)
价格与库存信息目前不能提供
Sigma-Aldrich
丙酮, puriss. p.a., ACS reagent, reag. ISO, reag. Ph. Eur., ≥99.5% (GC)
价格与库存信息目前不能提供
Sigma-Aldrich
无水碳酸钠, ACS reagent (primary standard), anhydrous, 99.95-100.05% dry basis
价格与库存信息目前不能提供
Sigma-Aldrich
甲醇, Laboratory Reagent, ≥99.6%
价格与库存信息目前不能提供
Sigma-Aldrich
丹磺酰氯, BioReagent, suitable for amino acid labeling, powder and chunks, ≥99% (HPLC)
价格与库存信息目前不能提供
Sigma-Aldrich
碳酸氢钠, anhydrous, free-flowing, Redi-Dri, ACS reagent, ≥99.7%
价格与库存信息目前不能提供
Sigma-Aldrich
无水碳酸钠, ReagentPlus®, ≥99.5%
价格与库存信息目前不能提供
Sigma-Aldrich
丙酮, suitable for HPLC, ≥99.8%
价格与库存信息目前不能提供
Sigma-Aldrich
乙酸铵, for molecular biology, ≥98%
价格与库存信息目前不能提供
Sigma-Aldrich
甲醇, ACS spectrophotometric grade, ≥99.9%
价格与库存信息目前不能提供