Porcine endogenous retrovirus (PERV) genomes are present in all pig cells. In this retrospective study, we assessed PERV infectivity in 28 patients treated with an extracorporeal bioartificial liver (HepatAssist system) that includes a membrane device containing porcine hepatocytes. All patients tested negative for PERV using polymerase chain reaction analysis of peripheral blood mononuclear cells (PBMC) collected up to 5 years after treatment. In vitro results showed that the membrane decreased the risk of PERV transmission by a factor of 10⁵, and porcine hepatocytes did not produce infectious PERV in co‐cultures with human cell line 293. Our results do not support the presence of PERV infection in patients treated with this porcine hepatocyte‐based bioartificial liver.