Porcine Hepatocytes: Isolation and Liver Tissue Engineering for Xenotransplantation.

Abstract

Various liver diseases result in liver failure, and liver transplantation as a definite treatment is limited by the shortage of organs available for transplantation. The use of isolated primary hepatocytes in cell-based therapies including hepatocyte transplantation, tissue engineering liver transplantation, and bioartificial liver support systems has gained increasing interest during the past years. Human hepatocytes are the preferred source of cells. Aside from the organ shortage, the isolation of human liver cells is usually limited by obtaining a sufficient quantity of high-quality, metabolically active cells. Furthermore, livers from which hepatocytes are typically harvested are not suitable for transplantation, with the variability in quantity and quality. Porcine hepatocytes, on the other hand, have the ability to perform complex biological functions and show modifiable behavior. Primary porcine hepatocytes are currently widely used in the investigation of drug metabolism, hepatotoxicity, protein biosynthesis, and gene expression. Primary hepatocytes do not proliferate in vitro and are sensitive to freeze-thaw damage in cryopreservation and thus need to be freshly isolated for each experiment. Consequently, the methods of porcine hepatocyte isolation are being actively sought after. Our laboratories have been involved in various applications of liver cells, and we have long-lasting experiences in liver cell isolation and their application in R&D. We here summarize the present protocol of our laboratories for primary hepatocyte isolation from pig and their liver tissue engineering for xenotransplantation.