Finding a Direct Method for a Dynamic Process: The DD (Direct and Dynamic) Cell-Tox Method

• Published in: International journal of molecular sciences 2024-05, Vol.25 (10), p.5133
• Main Authors: Madorran, Eneko, Kocbek Šaherl, Lidija, Rakuša, Mateja, Takač, Iztok, Munda, Miha
• Format: Article
• Language: English
• Subjects: Animals; Cell death; Cell division; Cell Survival - drug effects; cell viability; Cells, Cultured; Coculture Techniques - methods; Death & dying; dynamic direct cell death assay; Endothelial Cells - drug effects; Endothelial Cells - metabolism; Fatty acids; Fluorouracil - pharmacology; Hepatic Stellate Cells - drug effects; Hepatic Stellate Cells - metabolism; Hepatocytes - drug effects; Hepatocytes - metabolism; Humans; Hydrogen Peroxide - pharmacology; Ibuprofen - pharmacology; in vitro toxicology; Liver; Liver - cytology; Liver - drug effects; Liver - metabolism; Macrophages - drug effects; Macrophages - metabolism; Methods; Nonsteroidal anti-inflammatory drugs; Physiological aspects; Rifampin - pharmacology; Toxicity; Toxicity Tests - methods; Ultraviolet Rays
• ISSN: 1422-0067; 1661-6596; 1422-0067
• DOI: 10.3390/ijms25105133

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.