The Experimental Studies Of Hemo-perfusion With Primary Porcine Hepatocytes Immobilized In Hollow-fiber Module On Fulminant Hepatic Failure In Canine Models

Fulminant hepatic failure(FHF) is a severe diseases resulted in high mortality. At present, liver transplantation is an effective theatment, but the shortage of liver donors and using immunologic inhibitor long-term are a severe problem. Unfortunately, because of the shortage of liver donors ,a large number of patients with fulminant hepatic failure die before an organ becomes available for transplantation;some side effect from the use of immunologic inhibitor long-term will appear; some FHF patients may recover by themselves. There is, therefore, a need to develop a artificial liver support system for either providing adequate metabolic support to the remaining functional liver mass until it recovers from injury or serving as a "bridge" until a liver donor becomes available for transplantation. In this study , primary porcine hepatocytes were isolated and cultured . A bioartif icial liver support system was developed . A FHF dog model was made and performed in vivo experiment with the system. The efficacy and safety of BAL were assessed.Primary porcine hepatocytes were isolated by reformed collagenase perfusion technique . The average yield of porcine hepatocytes was1. 55xl010 cells per young pig with an average viability of 90. 2%?. 5% assessed by trypan blue exclusion. In the dynamic culture, hepatocytes had a tendency to aggregate and 85% of cells formed spheroids within 24h of culturing and were found to have viability and differentiating function during in vitro culture for 7 days(include albumin excretion, urea synthesis). The viability and differentiating function of hepatocytes were higher in the first three days Therefore they are an excellent source for bioartificial liver.The FHF canine model was made by the two-step devascularization method . The hepato-doudenal ligament was completely obstructed Ih after lateral-lateral porta-caval had been shunted for 48h. The dogs survived 30.5 + 20.48h. There was ideal consistency on the biochemical indexes(include albumin,ALT, AST, TBIL, DBIL, AM, PT, FIB, BCAA/AAA). The pathological examinations under light microscopy indicated hepatocytes necrosed and died of MSOF after fulminant hepatic failure.After isolated porcine hepatocytes were cultured and the FHF canine model was established, a bioreactor was prepared with cultured porcine hepatocytes and hollow-fiber module. Five FHF canine models were infused with BAL . Three FHF canine models were treated with empty-BAL. The experiment lasted for 16h.. The survival time of BAL group(96. 2?33.12h) was prolonged compared with the result in dogs with the empty-BAL group (37 + 9.93) (KO. 05). After 12h of treatment .Compared with the empty-BAL group and pre-treatment, the biochemical indexes of the BAL group such as TBIL, DBIL changed with statistical significance (P