There is growing interest in new therapeutic options for the treatment acute liver failure or to bridge liver function after extensive liver resections or small for size liver transplantation.We recently described a well differentiated human monocyte derived hepatocyte-like cell, which we can now generate form porcine monocytes.The porcine cells show similar function and are to be tested in an experimental model of extended liver resection, to bridge liver function.Methods: Monocytes were isolated from blood and after a six day treatment with MCSF and IL-3 were then exposed to a hepatocyte differentiation media containing FGF4 for 10 to 20 days.Neo-hepatocytes were analyzed for Albumin, Cytochrome P450 2D6 and 2C9, Connexin32, Actin and b-catenin.Urea formation was measured in vitro.Phase I biotransformation activity of neo-hepatocytes was verified using 7-ethoxycoumarin O-de-ethylase.The main phase II reactant, uridine di-phosphate (UDP)glucuronyltransferase was measured.1 x 107 human "neo-hepatocytes" were injected into the liver parenchyma of healthy mice to investigate albumin expression and integration into the liver parenchyma.Results: After 14-20 days cells displayed a typical hexagonal shape.Cells expressed, Cytochrome P450 2D6 and 2C9.Intermediate filaments were demonstrated by actin, intercellular connection characteristically by connexin 32.b-Catenin was expressed in some of the cells on the surface.Functional analysis of the neo-hepatocytes revealed a sustained albumin synthesis and a detoxifying capacity (urea production).Furthermore we found activity of 7-ethoxycoumarin O-deethylase in neohepatocytes.Phase II Metabolism was shown by UDP-glucuronidation.After injection of 1x107 Neo-hepatocytes we found integration into mice liver and excluded a fusion event.Cells expressed albumin.Our new findings indicate that porcine monocyte derived neo-hepatocytes have similar in vitro function with regards to synthesis and excretion aas well as detoxification and are now to be tested in a model of extended liver resection.
