ISOLATION AND CULTURE OF PORCINE HEPATOCYTES FOR ARTIFICIAL LIVER SUPPORT

The primary requirement of cells in a liver support system is the pres ervation of the in vivo metabolic functions that prevent or decrease t he progress of hepatic encephalopathy (HE) by providing interim suppor t to liver failure patients. While rodent hepatocytes offer a model fo r liver assist device (LAD) research, their limited number per animal prohibits direct scale up to human devices. Healthy human liver cells are seldom available in adequate numbers to support clinical LAD use; consequently, a large animal source of liver cells is needed. The stud y presented here explored the potential of porcine hepatocytes to prol iferate and maintain metabolic function in vitro. Porcine hepatocytes were isolated from similar to 12 kg swine by a modification of Seglen' s method. Hepatocytes cultured up to 10 days were shown to metabolize ammonia and maintain both Phase I and II detoxification functions. In addition, the cultures showed proliferative activity both as an increa se in total protein content and by thymidine incorporation. Immunocyto chemical staining identified cell proliferation through Day 4 to be pr imarily hepatocytes while Days 6 and 10 showed nonparenchymal cells to be increasing. The detoxification functions measured showed peak acti vity on Day 4 and gradually declined through Day 10. The ability of po rcine hepatocytes to proliferate and maintain a diversity of hepatic f unctions in culture strongly suggests their potential for use as the b iological component of artificial LADs.