Accumulated chemicals have been impairing water quality over decades of human activities. Organisms such as fish are often used for risk assessment, as chemical analysis is insufficient to assess the impact on health. Consequently, researchers have faced challenges in ecotoxicology, concerning the use of animals. Primary and cell line cultures are good alternatives for reducing animal use. This study aims to evaluate the use of primary hepatocytes culture from Oreochromis niloticus as an in vitro tool for screening environmental contaminants and pollutants. Metal cadmium (Cd) is the pollutant model.