The perfusion culture system of hepatocytes entrapped in a three-dimensional hollow fiber module with collagen gel is expected to realize an effective bioartificial liver (BAL) support system. The BAL module contained 5.4 billion porcine hepatocytes, which is approximately 7% of the total liver, supplied adequate metabolic functions, and improved the survival of the experimental acute liver failure swine. Improvement of the prothrombin time by the BAL treatment was not significant; however, the bleeding tendency was suppressed, which resulted in a significantly prolonged survival time of the animals in the BAL treatment group. A drastic up-regulation of the expression of the mRNA for plasminogen activator inhibitor 1 (PAI-1), which is known as an inhibitor for fibrinolysis, may explain the suppression of the bleeding tendency. These results support the efficacy of our BAL system for the treatment of acute liver failure.