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This limitation influences the cells response to drugs, potentially altering the drugs mechanisms of action, or boosting the cells resistance to the drug ( Spheroids, a 3D cell culture technique that closely mimics the cell status/environment within the liver, can produce results that more accurately reflect the in vivo changes in metabolic pathways in response to toxic compounds, as compared to 2D cell culture ( in vivo ( Research has shown that using 3D spheroid cultures of L-02 cells to investigate the hepatotoxicity of perfluorooctanoic acid (PFOA) through non-targeted metabolomics can provide valuable insights into the chronic toxicity mechanisms of PFOA, that cannot be uncovered in 2D monolayer models ( HLMs are capable of reactions catalyzed by membrane-bound enzymes such as CYP enzymes, FMOs, and uridine 5-diphospho-glucuronosyltransferases (UGTs) which are among the most relevant ones in drug metabolism

Ethics declarations Ethical approval All experiments were carried out in conformity with the European Union Directive (2010/63/EU), the Polish Act on the Protection of Animals (Dz.U

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