Isolation And Culture Of Porcine Hepatocytes And Manipulation Of AMP-Activated Protein Kinase

The experiment was conducted to study the effects of activator 5-amino-4-imidozolecarboxamide riboside (AICAR) and inhibitor Br8-AMP on AMPK of porcine hepatocytes and explore a method of isolation and culture of pig liver cells. Hepatocytes were isolated by two-step perfusion collagenase and cultured in suspension without gasing with O2/CO2(95 : 5) .The harvested cells were divided three treatments: the control, the activation and inhibiton incubated with 0.2mmol/l AICAR and'Br -AMP in medium respectively. The ALT,LDH ,BU of medium and the activity AMPK of hepatocytes were measured. The result showed:A number of hepatocytes with fine viability and specific function were harvested by the method of isolation and simplified culture in the experiment. The activity of AMPK were increased by 8.11% and 46.20% (p<0.01) respectively with AICAR at SOmin and 60min, Br8-AMP decreased the activity by 9.13%<, 22.39% respectively.The data indicateAICAR and Br -AMP can regulate effectively the activity of AMPK in vitro.