Relevance of the incubation period in cytotoxicity testing with primary human hepatocytes.
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Citations
Prediction of human drug-induced liver injury (DILI) in relation to oral doses and blood concentrations
Road Map for Development of Stem Cell-Based Alternative Test Methods.
Comparing in vitro human liver models to in vivo human liver using RNA-Seq
Setting the stage for next-generation risk assessment with non-animal approaches : the EU-ToxRisk project experience
Making Big Sense From Big Data.
References
Interspecies extrapolation by physiologically based pharmacokinetic modeling.
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Frequently Asked Questions (12)
Q2. What are the future works in "Relevance of the incubation period in cytotoxicity testing with primary human hepatocytes" ?
Therefore, 2 days may represent an adequate choice for cytotoxicity tests with human hepatocytes in future studies, offering the practical advantage that less culture medium changes are required. However, these results should be interpreted with caution and further reproduction is required before possible biological explanations are discussed.
Q3. Why was the function approximated according to the least square method?
Due to the non-linearity of the 4pLL model, the function was approximated according to the least square method with the Gauss–Newton algorithm.
Q4. How many cells were plated in 96-well plates?
After cell counting using Trypan blue to determine viability, 50,000 cells in FCS-containing medium were plated into each well of 96-well plates and kept at 37 °C for at least 3 h.
Q5. How long did the cells remain in the medium?
For single exposure, the cells were incubated with compounds for 24 h or 48 h; for repeated exposure, the compound-containing medium was renewed every 48 h and the cells were incubated for a total of 7 days.
Q6. How long does it take to wash out a test compound?
washout experiments with an initial exposure period with test compounds followed by test compound-free incubation or repeated exposure and washout periods may be considered.
Q7. What is the advantage of this procedure?
The advantage of this fitting procedure is that the left asymptote is used as a control level for calculation of EC50 and EC20 values which are more robust than just using the values of the solvent controls.
Q8. When was the solvent concentration increased to 0.5%?
Only when the cytotoxic test compound concentrations were not reached with 0.1% DMSO, the solvent concentration was increased to 0.5%.
Q9. What are the three compounds with a relatively strong decrease in EC50 after 7 days ?
Compounds with a relatively strong decrease in EC50 after 7 days compared to 1 day are busulfan, famotidine, and isoniazid (Fig. 2b).
Q10. What was the effect of the solvent on cell viability?
The applied solvent concentrations of 0.1% or 0.5% DMSO did not cause any cytotoxicity compared to cells cultivated in medium without DMSO.
Q11. What is the common cause of liver injury?
Dimethyl sulfoxide DILI Drug-induced liver injury EtOH Ethanol FAM Famotidine Glc Glucose HYZ Hydroxyzine INAH Isoniazid KC Ketoconazole LAB Labetalol LEV Levofloxacin MEL Melatonin MePa Methylparaben NAC N-Acetylcysteine NIM Nimesulide NFT Nitrofurantoin PhB Phenylbutazone PMZ Promethazine PPL Propranolol RIF Rifampicin TSN Triclosan VPA Valproic acid Vit C Vitamin CDrug-induced liver injury (DILI) is one of the principal reasons for drug withdrawal from the market (Godoy et al. 2013; Hewitt et al. 2007).
Q12. How long does it take to establish in vitro tests?
large efforts are undertaken to establish in vitro tests with the long-term goal to predict human toxicity (Daneshian et al.