Retinoic acid maintains differentiated cell morphology and functions in long-term cultured porcine hepatocytes: Obtaining functional cells for prolonged treatments with bioartificial liver

Porcine hepatocytes show several immunological characteristics and enzymatic activities of human liver, representing an ideal xenogenic source of cells as biological component of bioartificial liver (BAL). Isolated hepatocytes rapidly lose their specific metabolic activities and their typical morphology when cultured in the presence of serum. Since in BAL porcine hepatocytes are perfused by the patient's plasma, procedures able to minimize de-differentiation of cells could be useful for long-term treatment of acute liver failure (ALF). In this work we found that, in the presence of micromolar concentration of All trans-retinoic acid (ATRA), porcine parenchymal liver cells undergo to a lower extent the de-differentiating effects of long-term culture in the presence of serum. The evaluation of lidocaine metabolism showed that ATRA-treated cells retain specific hepatocyte function for a significantly longer time when compared to control hepatocytes. A tyrosine phosphorylation of PLC-yl was observed in concomitance with the ATRA-induced maximal functional activity. An increased expression of PLC-β3 and PKC-a and -β2 was also evidentiated at the longer time points explored, when the effects of ATRA in preservation of the differentiated morphology were maximal. These results provide the first evidence that ATRA plays a differentiating role in adult porcine hepatocytes cultured under de-differentiating conditions. The administration of ATRA to isolated parenchymal cells from pig liver may provide functional hepatocytes for prolonged treatment with BAL.