Background: Precision-cut liver slices (PCLS) preserve both the native tissue architecture and the multicellular interactions making them a valuable ex vivo model for studying hepatic physiology, drug metabolism and disease mechanisms. Nevertheless, during extended periods of cultivation this model is limited by a progressive reduction in tissue viability and function. Consequently, establishing a robust model that provides high viability, physiological relevance and native cell composition throughout mid- and long-term cultivation is crucial for expanding their application in regenerative medicine research.

Methods: Liver tissue from either healthy adult rats or pigs are recovered under sterile conditions and processed into 250 µm thick slices using a vibratome (Leica VT1200S). Slices are cultured for 5 days under optimized conditions in an incubator with a built-in rocking platform to improve oxygen/nutrient diffusion and waste removal. Culture media is supplemented with antioxidants and growth factors hypothesized to support prolonged tissue viability. Preliminary assessment includes metabolic activity assays, histological evaluation and lactate dehydrogenase (LDH) release.

Expected results: Preliminary data suggests that optimized processing and culture conditions improves tissue viability and functionality in both models. We expect to achieve sustained viability and metabolic activity for at least 7 to 10 days, with preserved histoarchitecture and reduced apoptotic signalling. We anticipate that this platform will be suitable for testing regenerative medicine approaches such as cell-derived hepatocyte transplantation and biomaterial scaffolding.

Conclusion: This model provides a versatile and translationally relevant platform for the study of liver repair processes and evaluation of emerging regenerative therapies such as cell-based treatments and bioengineered scaffolds. Importantly, this approach aligns with the 3R principle – reducing the number of animals, refining experimental methods and offering a partial replacement for in vivo studies in early-stage research.