Fig. 3

Cessation of immortalized hepatocyte-like cell (imHC) proliferation upon reaching confluency. The numbers of hepatocytes maintained in MEM/F12 medium containing 10% fetal bovine serum were recorded for more than 2 weeks to generate a standard growth curve. This experiment was performed in triplicate, and cell densities are presented as means ± standard deviations. imHCs exhibited a slower growth pattern than the widely used HC-04 hepatoma cell line (a) and stopped growing upon reaching confluency. Representative images of imHCs and HC-04 cells at days 1, 4, 8, 10, and 14 after subculture (b). HC-04 cells did not stop growing and formed multilayers of cells during a long-term culture, whereas imHCs maintained monolayer growth. Scale bar = 50 μm