PORCINE HEPATOCYTES FOR BIOHYBRID ARTIFICIAL LIVER DEVICES - A COMPARISON OF HYPOTHERMIC STORAGE TECHNIQUES

Two hypothermic preservation techniques were investigated to assess th eir possible role in on-demand cell supply for bioartificial liver sup port devices. Porcine hepatocytes from slaughterhouse organs were isol ated and either cold stored in a modified University of Wisconsin solu tion 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 subseque nt long-term culture (14 days) in both groups. Cold storage for 72 h r esulted in a decreased viability of cells (58.7 +/- 7.9%) with well pr eserved ultrastructures in the remainder of cells. In subsequent cultu re, albumin secretion was slightly increased, and cytochrome P450 IA1 dependent 7-ethoxycoumarine deethylation activity was reduced to about 40% of control values. After cryopreservation, hepatocyte cultures re vealed no severe damage to ultrastructures of cells, and functional pa rameters (albumin, 7-ethoxycoumarine deethylation) were comparable wit h controls after an initial drop in activity directly after thawing. L ength of storage time did not influence results. Both hypothermic pres ervation protocols might eventually play an important role for bioarti ficial liver processing and on-demand cell supply, dependent on the in dividual reactor design.