Bioartificial liver development using reversibly immortalized human liver cell lines.

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Acute liver failure (ALF) is often life-threatening and dramatically diminishes the quality of life of patients. Orthotopic liver transplantation has become a successful therapy of ALF, but this procedure is highly costly, limited by the scarcity of donor livers and associated with high morbidity and mortality. There is a compelling need for developing effective alternatives for patients with ALF. Considering the potential of the liver to regenerate, temporary support with bioartificial livers (BALs) is an attractive approach. Since technologies of tissue cell culture and biomaterials have been greatly advanced, many designs of BALs, including (1) a biological component, (2) a bioreactor, and (3) a whole blood or plasm a perfusion system are currently under investigating. It is unlikely that human hepatocytes can be isolated on a scale sufficient to treat more than a fraction of the patients who need bioartificial liver (BAL) treatment. The use of animal cells results in the concerns related to the transmission of infectious pathogens and immunologic and physiologic incompatibilities between the donor and humans. Human embryonic stem cells and bone marrow multipotent adult progenitor cells have receive great attention as a possible source for BALs. The use of tightly regulated clonal hepatocyte cell lines would be attractive. Such cell lines grow economically in tissue culture and provide the advantage of uniformity, sterility, and freedom of pathogens. In this project, we have made great efforts to develop BALs using reversibly immortalized human liver cells based on Cre/loxP-mediated site-specific recombination and achieved the following results :1) Enhancement of hepatic differentiated functions with p21 transduction,2) Establishment of a human liver endothelial cell line using a retroviral vector SSR#69 encoding a loxP-flanked simian virus 40 large T antigen cDNA,3) Establishment of hepatocyte isolation method from a surgically resected porcine hepatic segment.