The mini BioArtificiaLiver: a cellular biosensor in the drug development process B.Andria 1 , A. Bracco 1 , E.Alimenti 1 , C.Attanasio 1 , A.Tammaro 1 , S.Scala 1 , RAFM Chamuleau 3 , F.Calise 1&2 1 Center of Biotechnologies, 2 Liver Transplant Unit, Antonio Cardarelli Hospital 3 Academic Medical Center, Amsterdam Address: 9, Antonio Cardarelli Street – 80131 – Naples – Italy Tel & Fax: 0039 081 747 34 33; e-mail: biotecnologie@ospedalecardarelli.it Aims Every year a great amount of financial resources is spent by pharmaceutical companies on unsuccessful Clinical Trials because of poor activity/toxicity ratio, pharmacokinetics and pharmacodinamics accidents including poor Absorption-Distribution-Metabolism- Excretion/Toxicology profiles. Considering the key role accomplished by the liver in toxicology and metabolism of drugs there is a great need for screening tools using human cells. Furthermore the set up of bio-systems using human cells and able to test candidate drugs could be helpful in reducing the use of animals in the experimental testing as well as lowering costs concerning the drug development process. Methods In the Center of Biotechnologies of Cardarelli Hospital, in Naples, in collaboration with the Academic Medical Center of Amsterdam University, we assembled a mini Bioartificial Liver (mini BAL) able to host approximately 300,000 millions of viable human hepatocytes. The mini BAL is a three-dimensional system which resembles the best conditions for hepatocyte culturing and, differently from other monolayer approaches, shows a system-integrated oxygenation addressed to metabolism and respiratory functions optimization. Human hepatocytes are obtained by livers discarded from transplantation as well as surgical liver resections. Considering the low availability of human liver cells and in view of our experience, we used porcine hepatocytes in order to standardize the overall process. Previously we were able to successfully investigate in the mini BALs the effects of different doses of amphetamine, a well-known hepatotoxic drug, for 7 days. In this experiment we assessed the function of n° 3 minibal charged with 300,000 million viable human hepatocytes (viability≥ 80%) evaluating the specific cell functions through urea production, ammonia clearance, albumin synthesis. We observed a sharp, dose-dependent, decrease in hepatocytes functionality (urea production) with a restoration of the functionality in the following days.