STABLE HEPATOCYTE CULTURES FOR IN VITRO CARCINOGENESIS

THE OBJECTIVE IS TO ESTABLISH CULTURE CONDITIONS THAT WILL MAINTAIN HIGH LEVELS OF DIFFERENTIATED LIVER-SPECIFIC FUNCTIONS IN RAT AND HUMAN HEPATOCYTES FOR EXTENDED PERIODS OF TIME AS MEASURED BY THE MAINTENANCE OF NORMAL LEVELS OF METABOLIC FUNCTIONS. A FURTHER OBJECTIVE IS TO DEMONSTRATE THE UTILITY OF THE IMPROVED HEPATOCYTE CULTURES FOR IN VITROTOXICOLOGICAL, CARCINOGENIC, AND BIOCHEMICAL STUDIES AS A PREFERRED METHOD FOR THE INVESTIGATION OF TOXICOLOGICAL AND CARCINOGENIC MECHANISMS. STABLE EXTENDED CULTURE ALLOWS FOR ASSAY OF TISSUE-SPECIFIC FUNCTIONS AND THE OBSERVATIONS OF EVENTS THAT MAY BE CORRELATED WITH CARCINOGENIC ACTION. STABLE CULTURE HAS IMPORTANT PRACTICAL ADVANTAGES DUE TO THE AVAILABILITY OF CELLS FROM A SINGLE HUMAN DONOR FOR MANY TEST CAMPAIGNS AND THE ABILITY TO THOROUGHLY CHARACTERIZE THESE CULTURES. SUCH PRACTICAL ADVANTAGES WILL BE CRITICAL FOR COMMERCIALIZATION OF IN VITRO TOXICOLOGICAL TESTS. THE APPROACH UTILIZES A COLLAGEN-GLYCOSAMINOGLYCAN COPOLYMER CHEMISTRY THAT ALLOWS STABLE AND STERILE MEMBRANESTO BE CONSTRUCTED FROM PURIFIED COMPONENTS OF THE EXTRACELLULAR MATRIX. THESE MATERIALS TOGETHER WITH ASSAYS OF TISSUE-SPECIFIC FUNCTION WILL BE COMBINED INTO TEST KITS FOR TOXICOLOGICAL STUDIES.