A new type of mixed bioartificial liver (HBAL) was designed to evaluate its efficacy by simulating the purification of serum in liver failure and to explore the feasibility of its clinical application. Methods: Chinese human hepatocyte line -1 (CL-1) was selected as the donor of liver cells. In the microgravity environment, the hepatocyte microcarriers were co-cultured for 5 days, the total cells were about 4.0 × 109 cells, and the cell density was about 4.0 × 107 / ml. And then poured into a self-made bioreactor in a sterile environment to produce a biological part. Abiotic part of the use of blood perfusion + bilirubin adsorption. The biological and non-biological parts form a closed loop through two sets of polyethylene hoses. (CDB), cholic acid (CA) and blood ammonia (AA) in the serum of simulated liver failure were measured. The changes of the concentration of unbound bilirubin (UBD), chenodeoxycholic acid, cholic acid (CA) and blood ammonia (AA) Changes of hepatocyte function, morphology and liver cell activity. Results: The concentrations of UBD, chenodeoxycholic acid, CA and AA in the serum of simulated liver failure were significantly higher than those in the control group (335.3 ± 6.0) μmol / L, (395.0 ± 5.6) μmol / L, (155.7 ± 5) (41.8 ± 28.7) μmol / L, (42.2 ± 7.3) μmol / L, (3.5 ± 1.0) μmol / L, (0.3.0 ± 28.7) μmol / L, (P <0.05). After 24 h, the concentration of CL-1 cells changed after 48 h of circulation. The serum of C-group was significantly higher than that in control group (P <0.05), and the difference was statistically significant (P <0.05) (25.9 ± 4.2) IU / L, (22.0 ± 3.6) IU / L, (0.28 ± 0.09) μmol / L), and the activity of aspartate aminotransferase (ALT), aspartate aminotransferase (AST) and lactate dehydrogenase (LDH) (31.0 ± 2.6) IU / L, (31.6 ± 8.0) IU / L, (0.41 ± 0.12) μmol / L, and the difference was statistically significant (P <0.05). The number and viability of hepatocytes in the reactor were also significantly decreased after 48 h (P <0.05). Conclusion: (1) We constructed a new type of perfusion bioreactor. CL-1 cells in this bioreactor can maintain high activity and good function. (2) The new mixed biological human liver can significantly reduce the concentration of UBD, chenodeoxycholic acid, CA and AA in serum of simulated liver failure, and has the function of removing these toxic substances, which indicates that it has obvious liver function support.