Wiström, Linda2025-06-232025-06-232025-06-23https://hdl.handle.net/2077/88142Animal-derived products, such as fetal bovine serum and porcine derived trypsin, has been utilized within cell cultivations for decades. However, these animal-derived products come with an increased environmental impact, technical difficulties and animal welfare issues, where the scientific community has expressed a need for an animal-product-free cell culture protocol. The aim of this study was therefore to evaluate the applicability of a recently established animal product-free protocol for mammalian cell cultures on the Poeciliopsis Lucida Hepatocellular Carcinoma (PLHC-1) fish cell line, in the context of the environmental toxicity assay ethoxyresorufin-O-deethylase (EROD) activity. The aim was answered by cultivating PLHC-1 according to the animal-product-free cell culture protocol and compare growth, vitality and EROD activity to cells cultured with the standard animal-derived protocol. This study found that animal free CTS™ TrypLE™ Select Enzyme did not have a negative effect on cell vitality or estimated EROD activity, making it a comparable option to porcine derived trypsin-EDTA (0.05 %). However, the results also displayed that PLHC-1 could not be cultured long-term or display measurable levels of EROD activity when cultured fully according to the animal-product-free protocol. These findings conclude that trypsin-EDTA has the possibility to be replaced by CTS™ TrypLE™ Select Enzyme, but further optimization of the animal-free-protocol is needed to replace fetal bovine serum. The substitution to animal-free CTS™ TrypLE™ Select Enzyme could lower the environmental impact of the animal-derived protocol typically used for PLHC-1 and the environmental toxicity assay EROD activity as well as aligning the protocol closer to animal welfare principles.engPLHC-1, djurfritt cellodlingsprotokoll, EROD, CYP1A, fosterbovint serum, β-naftoflavon, trypsin, animal-free cell culture protocol, fetal bovine serumText