In vitro methodologies for enhanced toxicity testing

DelRaso, Nicholas J.

doi:10.1016/0378-4274(93)90122-e

Cited by 26 publications

(18 citation statements)

References 16 publications

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“…1, 2 The use of in vitro model systems in toxicity testing has many advantages including the decrease in animal numbers, the reduced cost of animal maintenance and care, small quantity of a chemical needed for testing, shortening of the time needed, and increase in throughput for evaluating multiple chemicals and their metabolites. 3, 4 In vitro systems also allow to study chemical metabolism, evaluate the mechanisms of toxicity, measure enzyme kinetics, and examine dose-response relationships. 4 …”

Section: Chemical Toxicity Testingmentioning

confidence: 99%

In vitro models for liver toxicity testing

et al. 2013

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Over the years, various liver-derived in vitro model systems have been developed to enable investigation of the potential adverse effects of chemicals and drugs. Liver tissue slices, isolated microsomes, perfused liver, immortalized cell lines, and primary hepatocytes have been used extensively. Immortalized cell lines and primary isolated liver cells are currently most widely used in vitro models for liver toxicity testing. Limited throughput, loss of viability, and decreases in liver-specific functionality and gene expression are common shortcomings of these models. Recent developments in the field of in vitro hepatotoxicity include three-dimensional tissue constructs and bioartificial livers, co-cultures of various cell types with hepatocytes, and differentiation of stem cells into hepatic lineage-like cells. In an attempt to provide a more physiological environment for cultured liver cells, some of the novel cell culture systems incorporate fluid flow, micro-circulation, and other forms of organotypic microenvironments. Co-cultures aim to preserve liver-specific morphology and functionality beyond those provided by cultures of pure parenchymal cells. Stem cells, both embryonic- and adult tissue-derived, may provide a limitless supply of hepatocytes from multiple individuals to improve reproducibility and enable testing of the individual-specific toxicity. This review describes various traditional and novel in vitro liver models and provides a perspective on the challenges and opportunities afforded by each individual test system.

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Section: Chemical Toxicity Testingmentioning

confidence: 99%

In vitro models for liver toxicity testing

et al. 2013

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“…In vitro systems are useful for dissecting cellular biochemical pathways of toxicity because the target cell can be studied directly without interference of other cell types or organ systems. 23 The human fetal hepatic cell line WRL-68 retains many liver-specific functions such as secretion of albumin, a cytokeratin pattern similar to hepatocytes, and characteristic enzymatic activities like GGT, ALAT and ASAT. 24 The purpose of the present investigation was to evaluate whether this cell line could be used as a model for the study of the effects that ethanol per se produces at the cellular level and to determine if these effects are similar to those described in in vivo systems.…”

Section: Introductionmentioning

confidence: 99%

The effect of chronic and acute ethanol treatment on morphology, lipid peroxidation, enzyme activities and Na+ transport systems on WRL-68 cells

et al. 1995

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In this study we measured some parameters that are asso ciated with ethanol damage to the liver. The method allowed us to determine the injury that chronic and acute ethanol treatments produce at the cellular level without interference from homeostatic or compensatory mecha nisms. The system used is a hepatic fetal human cell line, WRL-68, which retains, in culture, many of the liver-spe cific functions. WRL-68 cells do not metabolise ethanol, and consequently we could evaluate the effect of ethanol alone. We explored two different conditions: 30 days with 0.1 M ethanol (chronic treatment) and 24 h in the presence of 0.5 M ethanol (acute treatment). 1 The transmission electron microscopy studies revealed, in both treatments, the presence of granules not usually present in the cytoplasm of control cells and morphologi cal mitochondrial alterations in chronically treated cells. 2 Lipid peroxidation, measured as the rate of malondi aldehyde production, increased three and a half times in acutely treated cells and about twofold in chronically treated cells. 3 The percentage of total activity (activity in the medi um/(activity in the medium + activity of the cells). 100) and the enzymatic activity in the culture medium of gamma glutamyl transpeptidase (GGT), alanine amino transferase (ALAT), aspartate amino transferase (ASAT) and alkaline phosphatase (AI-P), increased. 4 We measured some parameters related to the transport of sodium across the membrane. Cells chronically treated with ethanol had higher rate constants and effluxes than control cells. There was no difference between the total and passive efflux. Ethanol treated cells apparently lacked the ouabain sensitive pathway. In acutely treated cells, the total sodium efflux and the rate constant were enhanced. Sodium pools in the acutely treated cells were diminished and active sodium pumping was seven times higher than in control cells. 5 We determined the number of high affinity ouabain binding sites per cell. Ethanol did not alter the number of pumps, rather it seems to induce a functional alteration. Our results indicate that ethanol per se induces lipid per oxidation, alters enzymatic activities, sodium transport systems, sodium pools and cellular morphology, and that all these changes may be partly responsible for ethanol- induced hepatotoxicity. The data compare favourably with those reported in the literature for many different systems. Therefore our model for studying the mechanism of alco hol effects appears to be valid, with the advantage of being able to compare experiments that can be done in the same system and under the same conditions.

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“…During the past decade, use of cell culture techniques to evaluate the toxicity of compounds in toxicological research have been developed (DelRaso 1993). Manual cell counting is time consuming, inaccurate, and unfeasible for adhering cell lines.…”

Section: Discussionmentioning

confidence: 99%

Comparative Measurement ofIn VitroParaquat and Aflatoxin B1 Cytotoxicity Using Three Different Cytotoxicity Assays in Pheochromocytoma Cells (PC-12)

Mohammadi-Bardbori

Nejati

Esmaeili

et al. 2008

Toxicology Mechanisms and Methods

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Among the herbicides and mycotoxins, paraquat (PQ) and aflatoxin B1 (AFB1) are highly cytotoxic. In this study the toxicity of PQ and AFB1 in the cultured cell were determined using MTT [3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide], JG-B (Janus green B), and NR (neutral red) assay by multiwell scanning spectophotometry. JG-B was used not only for the vital staining of mitochondria but also for viability assay and was compared to MTT and NR assay. Various concentrations of paraquat (0.1 mM to 100 mM) and AFB1 (0.001 nM to 10 nM) on the PC-12 cells were investigated. The 50% lethal concentration of toxins (LC50) were determined for PQ (7.70 +/- 2.50, 3.67 +/- 1.53, 4.85 +/- 2.44) and AFB1 (0.16 +/- 0.01, 0.13 +/- 0.04, 0.14 +/- 0.02) as determined by these methods (JG-B, NR, and MTT, respectively). A significant correlation was found among the JG-B and MTT using PQ (r(2) = 0.99, p < 0.05) and significant correlation was also found among the three methods (r(2) = 0.95, 0.93, and 0.92, p < 0.05) using AFB1. No significant correlation was found between JG-B and MTT with NR (r(2) = 0.34 and 0.35, p < 0.05, respectively) using PQ. These results suggest that both methods (MTT assay and JG-B assay) are reliable and are comparable for determining the cytotoxicity. It is concluded that the JG-B assay may be preferable to MTT assay methods because of its simplicity, low cost, sensitivity, and objectivity; in addition, this method takes little time to be done.

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In vitro methodologies for enhanced toxicity testing

Cited by 26 publications

References 16 publications

In vitro models for liver toxicity testing

In vitro models for liver toxicity testing

The effect of chronic and acute ethanol treatment on morphology, lipid peroxidation, enzyme activities and Na+ transport systems on WRL-68 cells

Comparative Measurement ofIn VitroParaquat and Aflatoxin B1 Cytotoxicity Using Three Different Cytotoxicity Assays in Pheochromocytoma Cells (PC-12)

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