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Published on: June 15, 2022
Finding a Direct Method for a Dynamic Process: The DD (Direct and Dynamic) Cell-Tox Method
Eneko Madorran1, Lidija Kocbek Šaherl1, Mateja Rakuša1
1Faculty of Medicine, Institute of Anatomy, Histology and Embryology, University of Maribor, Taborska Ulica 8, 2000 Maribor, Slovenia.
Abstract:
The main focus of in vitro toxicity assessment methods is to assess the viability of the cells, which is usually based on metabolism changes. Yet, when exposed to toxic substances, the cell triggers multiple signals in response. With this in mind, we have developed a promising cell-based toxicity method that observes various cell responses when exposed to toxic substances (either death, division, or remain viable). Based on the collective cell response, we observed and predicted the dynamics of the cell population to determine the toxicity of the toxicant. The method was tested with two different conformations: In the first conformation, we exposed a monoculture model of blood macrophages to UV light, hydrogen peroxide, nutrient deprivation, tetrabromobisphenol A, fatty acids, and 5-fluorouracil. In the second, we exposed a coculture liver model consisting of hepatocytes, hepatic stellate cells, Kupffer cells, and liver sinusoidal endothelial cells to rifampicin, ibuprofen, and 5-fluorouracil. The method showed good accuracy compared to established toxicity assessment methods. In addition, this approach provided more representative information on the toxic effects of the compounds, as it considers the different cellular responses induced by toxic agents.
Insights
This study introduces a novel cell-based toxicity assessment method that monitors diverse cellular responses beyond metabolism. The approach accurately predicts toxicant effects by analyzing cell population dynamics, offering more representative toxicity data.
Area of Science:
- Toxicology
- Cell Biology
- Biochemistry
Background:
- In vitro toxicity testing traditionally relies on metabolic activity to assess cell viability.
- Cells exhibit multifaceted responses to toxic substances, including changes in viability, division, and death.
- Existing methods may not fully capture the complexity of cellular reactions to toxicants.
Purpose of the Study:
- To develop and validate a novel cell-based toxicity assessment method.
- To evaluate diverse cellular responses (viability, division, death) to toxicants.
- To predict toxicant effects based on collective cell population dynamics.
Main Methods:
- Developed a cell-based assay observing multiple cellular responses to toxic substances.
- Tested the method using a monoculture of blood macrophages exposed to various agents (UV light, H2O2, nutrient deprivation, etc.).
- Validated the method with a coculture liver model (hepatocytes, stellate cells, Kupffer cells, endothelial cells) exposed to drugs (rifampicin, ibuprofen, 5-FU).
Main Results:
- The novel method demonstrated good accuracy when compared to established toxicity assessment techniques.
- The approach provided more comprehensive and representative information on toxic effects.
- Observed and predicted cell population dynamics based on collective cellular responses.
Conclusions:
- The developed cell-based toxicity method offers a more representative assessment of toxicant effects.
- Monitoring diverse cellular responses enhances the accuracy and information content of in vitro toxicity testing.
- This approach advances the field of in vitro toxicology by considering a broader spectrum of cellular reactions.
