Abstract Background A great many patients awaiting liver transplantation die because of the shortage of donor livers. To resolve the problem, liver support systems like bioartificial livers (BALs) have become subjects of active investigation. However, the problem with BALs is that it is difficult to find a source of healthy hepatic cells with good liver function. This study explored the possibility of employing Chang liver cells (ATCC CCL-13), a human hepatoma cell line as a source for liver support. Methods To evaluate the function of Chang liver cells in vitro, hepatocyte markers were measured by Western blotting and laser confocal microscopy. The gene expression of hepatic markers was examined by reverse transcriptase polymerase chain reaction (RT-PCR). After acute liver failure (ALF) was established by 90% partial hepatectomy, Chang liver cells were intrasplenically transplanted for treatment. Results In vitro, Western blotting and laser confocal microscopy showed conspicuous expression of liver function markers, such as albumin, uridine diphosphate glucuronosyltransferase, and cytochrome P450 3A4 by Chang liver cells. RT-PCR revealed expression of related genes at the mRNA level. The survival of rats receiving transplanted Chang liver cells reached 40% versus 0% among the controls (P < .01). Liver function of rats receiving transplanted Chang liver cells was improved at 24 hours after ALF, as evidenced by decreased levels of alanine transaminase, aspartate aminotransferase, bilirubin, and alkaline phosphate. Conclusions Chang liver cells, which express liver function markers and exert obvious liver-protective effects in ALF can serve in liver support systems.