2017 Volume 22 Issue 1 Pages 101-106
The development of a cell culture maintenance system is important for alternatives to animal testing and experimentation for drug discovery. Generally, cryopreserved human hepatocyte viability and function are maintained for approximately 3 days after thawing in two-dimensional (2D) monolayer culture using a collagen-coated dish. In this study, we cultured human hepatocytes in the silicate fiber-based three-dimensional (3D) scaffold Cellbed®. As a result of observation with a scanning electron microscope (SEM), hepatocytes were cuboidal as compared with monolayer culture. This cell morphology seems to be appropriate for maintenance of cell viability and function of hepatocytes. Cell viability in Cellbed culture was maintained at a higher level and for a longer period (about 2 weeks) than that of monolayer culture. Levels of glutamic oxaloacetic transaminase (GOT) and glutamic pyruvic transaminase (GPT) leakage were also lower in Cellbed culture than in monolayer culture. Based on gene expression analysis, CYP2C19 and CYP3A4 were expressed in Cellbed culture at levels more than 3.0 times higher than those of monolayer culture. Moreover, SLC22A1, the gene for bile acid transporter, was expressed in Cellbed culture at a level more than 2.9 times higher than that in monolayer. This culture system may be applied to hepatic metabolism studies and long-term in vitro liver toxicity testing.
