Porcine hepatocytes for biohybrid artificial liver devices: a comparison of hypothermic storage techniques

Two hypothermic preservation techniques were investigated to assess their possible role in on-demand cell supply for bioartificial liver support devices. Porcine hepatocytes from slaughterhouse organs were isolated and either cold stored in a modified University of Wisconsin solution for up to 72 h or directly cultured in a sandwich configuration, frozen at Day 3 of culture, and stored for up to 30 days with subsequent long-term culture (14 days) in both groups. Cold storage for 72 h resulted in a decreased viability of cells (58.7 +/- 7.9%) with well preserved ultrastructures in the remainder of cells. In subsequent culture, albumin secretion was slightly increased, and cytochrome P450 IA1 dependent 7-ethoxy-coumarine deethylation activity was reduced to about 40% of control values. After cryopreservation, hepatocyte cultures revealed no severe damage to ultrastructures of cells, and functional parameters (albumin, 7-ethoxycoumarine deethylation) were comparable with controls after an initial drop in activity directly after thawing. Length of storage time did not influence results. Both hypothermic preservation protocols might eventually play an important role for bioartificial liver processing and on-demand cell supply, dependent on the individual reactor design.