Effect of Low Temperature Preservation and Cell Density on Metabolic Function in a Bioartificial Live

Difficulties associated with bioartificial liver (BAL) preservation limit not only the commercialization of BAL, but also its clinical trials. In this study, the possibility of cold preservation of BAL cartridges containing porcine hepatocytes was examined at 4$^{\circ}C$. In an in vitro perfusion culture System, BAL cartridges maintained cytochrome P450 metabolic function for at least 50 days. However, all BAL cartridges completely lost their ammonia eliminating ability when stored at 4$^{\circ}C$. We a1so studied the effect of cell density on the maintenance of BAL liver function In a highly differentiated and healthy state. As expected, BALs containing a larger number of hepatocytes demonstrated higher metabolic functions. When metabolic functions were compared per gram of hepatotytes, no large differences were observed between devices containing different densities of hepatocytes. Decreased cell density did not Successfully prolong BAL function. The viability and function of isolated hepatotytes highly depend on the culture conditions, such as cell density, substrata, culture media, and additives to the culture media. Perfusion culture of BAL cartridges at 4$^{\circ}C$ gave a promosing result with respect to the maintenance of P450 activity. However, as indicated by the rapid loss of ammonia metabolic activity, many factors still remain to be optimized for preservation of BAL keeping high metabolic functions for a longer time.