AIM To develop a culture mode providingdurable biomaterials with high yields andactivities used in bioartificial liver.METHODS Hepatocytes were isolated from awhole pig liver by Seglen’s method of orthotopicperfusion with collagenase.In culture onmicrocarriers,primary porcine hepatocyteswere inoculated at a concentration of 5×10~7/mLinto the static culture systems containing 2 g/LCytodex-3,then supplemented with 100 mL/Lfetal calf serum(FCS)or 100 mL/L porcineportal vein serum(PPVS)respectively.Inspheroidal aggregate culture hepatocytes wereinoculated into 100 mL siliconized flasks at aconcentration of 5.0×10~6/mL.RESULTS In culture on microcarriershepatocytes tended to aggregate on Cytodex-3obviously after being inoculated.Typical multi-cellular aggregated spheroids could be found inthe two systems 24 h-48 h after hepatocyteswere cultured.The morphological charact-eristics and synthetic functions were maintainedfor 5 wk in FCS culture system and 8 wk in PPVSculture system.In spheroidal aggregate cultureabout 80%-90% isolated hepatocytes becameaggregated spheroids 24h after cultured insuspension and mean diameter of the spheroidswas 100μm.The relationship among thehepatocytes resembled that in the liver in vivo.Synthetic functions of albumin and urea of the spheroids were twice those of hepatocytescultured on monolayers.CONCLUSION As high-yields and high-activitymodes of culture on microcarriers or inspheroidal aggregate culture with portal veinserum are promising to provide biomaterials forbioartificial liver(BAL)efficiently. AIM To develop a culture mode providingrable biomaterials with high yields and activities used in bioartificial liver. METHODS Hepatocytes were isolated from awhole pig liver by Seglen’s method of orthotopicperfusion with collagenase. In culture on microarriers, primary porcine hepatocyteswere inoculated at a concentration of 5 × 10-7 / mLinto the static culture systems containing 2 g / LCytodex-3, then supplemented with 100 mL / L calal serum (FCS) or 100 mL / L porcineportal vein serum (PPVS) respectively. Inspheroidal aggregate culture hepatocytes were inoculated into 100 mL siliconized flasks at aconcentration of 5.0 × 10 6 / mL. RESULTS In culture on microcarriers of cells tended to aggregate on Cytodex-3obviously after being inoculated.Typical multi-cellular aggregated spheroids could be found inthe two systems 24 h-48 h after hepatocyteswere cultured. The morphological charact -eristics and synthetic functions were maintained for 5 wk in FCS culture system and 8 wk in PPVSculture system. In spheroi dal aggregate cultureabout 80% -90% isolated hepatocytes became aggated spheroids 24h after cultured insuspension and mean diameter of the spheroidswas 100 μm. The relationship among the hepatic cells resembled that in the liver in vivo. Synthetic functions of albumin and urea of ​​the spheroids were twice those of hepatocytes cultured on monolayers. CONCLUSION As high-yields and high-activity motes of culture on microcarriers or inspheroidal aggregate culture with portal vein serum are promising to provide biomaterials for bioartificial liver (BAL) efficiently.