Double-Layered Microphysiological Systems Improve Cell Adhesion, Viability, and Mitochondrial Activity in Human Hepatocyte Culture

Abstract

Microphysiological systems (MPS) provide a basis for culturing human hepatocytes and mimicking their function in viro. However, human hepatocytes do not proliferate and must be seeded at a high density. In a closed system device, such as the MPS, the recommended seeding density does not allow for good adhesion. In this study, we demonstrated that the perfusion of lower channel immediately after seeding in a double-layered MPS, Fluid3D-X^®, which has two-flow channels, improved cell adhesion and mitochondrial activity and enabled the healthy culture of hepatocytes, even when seeded at the recommended seeding density. Furthermore, in Fluid3D-X^®, which has a long flow channel, increasing the medium flow rate in the upper and lower two channels maintains cell viability throughout the entire channel, suggesting that even human hepatocytes with high oxygen demand can be cultured in a healthy manner. The approach developed in this study is expected to guide the broader application of MPS devices in studies of human hepatocytes.