Background: Three-dimensional (3D) cell culture techniques have gained considerable importance in recent years, especially in the field of liver research. Cultivating hepatocytes in two-dimensional (2D) environments is limited due to their rapid loss of functional properties and inability to maintain their native 3D architecture. In contrast, 3D cell cultures, including precision-cut liver slices (PCLS), aim to mimic cell-cell as well as cell-matrix interactions and thus in vivo liver conditions. This is critical for the study of liver function, toxicity testing, and the development of liver models for transplantation medicine. Due to the limited number of human organs available for transplantation research, porcine organs are commonly used for proof-of-concept studies, thus porcine hepatocytes serve as an excellent model for preliminary research.

Methods: Various 3D culture techniques, including spheroid cultures, hydrogel-based systems, and precision-cut liver slices, are used to investigate primary porcine hepatocytes. Hepatocytes from porcine liver biopsies are cultured in both 2D and 3D environments and PCLS are prepared by cutting liver tissue into thin, viable slices using a precision-cutting technique. Morphological and functional analyses are performed to assess cell viability, enzyme activities, and gene expression.

Expected Results: Hepatocytes in 3D cultures, including PCLS, are expected to exhibit higher activity, cell viability, and functional stability over an extended period of time compared to 2D cultures. Specifically, PCLS are expected to maintain a more physiological liver architecture and cell-cell interactions, providing a more accurate representation of in vivo liver conditions.

Conclusion: In conclusion, these experiments seek to demonstrate the efficacy of 3D cell culture methods, including PCLS, in preserving the physiological and functional characteristics of hepatocytes in vitro. These advanced culture systems provide valuable tools for liver research. Future studies should focus on further optimization and standardization of 3D culture techniques to enhance their application in biomedical research and translation into clinical practice.