Enhanced self-organization of size-controlled hepatocyte aggregates on oxygen permeable honeycomb microwell sheets

Three-dimensional hepatocyte aggregates possessing highly-metabolic functions are required for analytical applications, such as in drug discovery and drug toxicity screening, to obtain physiologically relevant outcomes. Here, we report a rapid and highly productive method to obtain size-controlled rat hepatocyte aggregates using oxygen permeable honeycomb microwells. Rapid cell aggregation occurred in honeycomb microwells even with increased cell density per unit area (four times higher than the cell density in a conventional polystyrene culture dish). The activity of cytochrome P450 (CYP) 1A1 and CYP3A2 was enhanced in the hepatocyte aggregates, 52–88 μ m in diameter, compared with those in the hepatocyte monolayer. These findings are expected to contribute to the rapid organization of optimal-sized hepatocyte aggregates for cell-based assays.