Microfluidic Perfusion Culture of Human Hepatocytes

Abstract

Analysis using human cells has been widely used in place of animal experiments. To obtain culture environments closer to those with in vivo, perfusion culture using microfluidic devices is being studied instead of stationary culture such as in a culture dish. With conventional perfusion culture with microfluidic devices, pumping system is externally provided, causing a large dead volume of culture medium. As a result, applied drugs as well as metabolites and signal transmitters from cells are diluted. We minimized this dead volume by embedding micropumps within the device to realize a high concentration of metabolites and signal transmitters from cells by perfusion with small amounts of culture medium and its effects on the cells. Using Hep-G2, established from a human hepatoma, we successfully formed Hep-G2 spheroids which are not observed in conventional culture. Evaluating activity from the DNA amount and albumin produced, we found that Hep-G2 spheroids formed in our device showed higher activity than conventional 2-dimensional culture. We demonstrated that the functionally highly integrated on-chip perfusion cell culture microdevice provided cells with a culture environment close to that in vivo and promoted morphological change and expression of high activity in cells.