Background: Ex vivo liver perfusion has emerged in the last years as a promising tool to maintain organ functions and complexity. Hence, it is not only of high importance for transplantation research but also as device or drug testing model. Here, we investigated the possible use of slaughterhouse material to develop a normothermic ex vivo liver perfusion model with an animal friendly approach. The strict processes in a slaughterhouse present a challenging environment for organ procurement and a critical starting state of the organ. Methods: Livers from two slaughterhouses (group 1 and 2) with different slaughtering procedures were compared to lab animal livers. Our developed ex vivo platform was used to reperfuse the livers with physiological pressures at 39°C for 5h. To assess functionality an ICG assay was performed. Further, blood gases, bile production, and gene expression were analyzed to test for viability and damage markers. Results: Physiological flow rates were achieved in all livers. Significant decreases were seen in bile production and functionality for group 1. However, ICG functionality test, bile production, and viability analyses showed comparable results for group 2 to livers from lab animals. Conclusion: This study showed the applicability of slaughterhouse material but indicated an important impact of different slaughter methods on liver performance. Further research will focus on minimization of harmful factors in the slaughterhouse and reconditioning livers before reperfusion, to enable a consistent model independent of slaughterhouse availability. This model allows device testing, organoid implantation and validation of new artificial organs with an animal friendly approach.