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Preserved liver-specific functions of hepatocytes in 3D co-culture with endothelial cell sheets.
Hepatocyte-based tissue engineering is an attractive method that is being developed to treat liver diseases. However, this method is limited by the relatively short lifespan of cultured hepatocytes to maintain their normal function. For this reason, the present study was designed to develop a cell sheet-based hepatocyte co-culture system that enables cultured hepatocytes to preserve their functions for a longer period of time. To achieve this goal, a monolayer cell sheet composed of endothelial cells (EC) was placed on top of a monolayer of hepatocytes (Hep). In this hybrid cell sheet format, histological examination revealed that bile canaliculi networks were formed and well developed among the hepatocytes in the layered Hep-EC sheet group. The albumin secretion level was highly preserved at least for 28 days in the hybrid Hep-EC sheet, whereas the monolayer of hepatocytes exhibited a markedly reduced time course of secretion. The expression levels of hepatocyte-specific genes including albumin, hepatocyte nucleus factor 4 (HNF 4), multidrug resistance-associated protein 2 (MRP 2), and claudin-3 were significantly higher in the Hep-EC sheet compared to the Hep sheet alone after 14-days in culture. In all, this culture system provides a valuable technology to prolong hepatocyte functionality and enable more efficient development of liver tissue engineering approaches to create liver-targeted regenerative therapies.
Institute of Advanced Biomedical Engineering and Science, Tokyo Women's Medical University, Tokyo, Japan., , ,
Fluorescent Antibody Technique
Gene Expression Regulation
Pub Type(s)Journal Article
Research Support, Non-U.S. Gov't