Biofabrication of Layered Membrane Systems by Using Human Hepatocytes and Endothelial Cells: A Comparative Study

Strategies to overcome in vitro dedifferentiation of hepatocytes preserving key features of the liver phenotype in vitro constitute a major goal in medical applications. The realization of biohybrid membrane systems, utilizing primary human hepatocytes in co-culture with non-parenchymal cells and polymeric membranes with suitable properties, offered interesting opportunities for the design of bioartificial liver. This study was aimed to investigate the morphological and functional behavior of primary human hepatocytes with endothelial cells (ECs) in two different layered co-culture membrane systems, which were realized by using a synthetic polymeric membrane and two different sequential cell seeding methods. We evaluated the heterotypic cell-cell interactions in the different layered membrane systems in order to understand if the variation of the heterotypic interface between hepatocytes and ECs and of cell type interacting with membrane may influence the performance of functional biofabricated hepatocellular constructs. Overall results demonstrated a strong influence of the heterotypic interactions on the performance of the tissue constructs in terms of albumin secretion, urea synthesis and drug biotransformation functions.