High yield hepatocyte isolation from pig livers for investigation of hybrid liver support systems: influence of collagenase concentration and body weight.

Cultured hepatocytes would be required in large quantities if a hybrid liver support system became available. We have therefore performed a study of larger scale enzymatic hepatocyte isolation on whole pig livers. A new method of open-loop and recirculating perfusion in five steps using both the portal venous and the hepatic arterial vascular systems has been developed.

Comparison of four methods for mass hepatocyte isolation from pig and human livers.

Using the pig liver, parameters for large scale hepatocyte isolation were studied in order to develop a technique suitable for human organs. These investigations led to a 5-step modification of the original 2-step method. Four groups were compared.

Comparison of pig hepatocyte isolation using intraoperative perfusion without warm ischemia and isolation of cells from abattoir organs after warm ischemia.

Enzymatic hepatocyte isolation using warm ischemic pig livers from an abattoir was compared with isolation using in situ perfused organs. Using organs from animals of 30 kg body mass (BM), the intraoperative perfusion showed superior results. The use of livers from abattoir pigs of 40-50 kg BM after warm ischemia resulted in a lower yield of hepatocytes and in high rates of injured cells.

Nonenzymatic versus enzymatic hepatocyte isolation from pig livers for larger scale investigations of liver cell perfusion systems.

A comparison of nonenzymatic and enzymatic hepatocyte isolation was performed on pig livers. The collagenase perfusion showed superior results: mean viability 72 +/- 10% versus a maximum viability of 21% using EDTA-perfusion. A five-step collagenase perfusion technique, developed for pig livers enables larger scale investigations, in order to develop methods for hepatocyte cultures in therapeutical liver cell perfusion systems.