A TECHNIQUE FOR PORCINE HEPATOCYTE HARVEST AND DESCRIPTION OF DIFFERENTIATED METABOLIC FUNCTIONS IN STATIC CULTURE

Current bioartificial liver devices are based on the use of a large ma ss of hepatocytes exhibiting differentiated metabolic function. The pi g has become a source of interest for the acquisition of such cells-ho wever, harvesting a large mass of highly viable cells has met with dif ficulty. This study describes a technique for harvesting large quantit ies of hepatocytes at viabilities greater than 90% and also describes several features documenting differentiated function. Pigs, 6 to 10 kg body weight, underwent in situ two-step whole liver perfusion (ethyle ne glycol tetraacetic acid and collagenase) and ex vivo cell harvest. Harvests yielded an average of 19.5 billion cells with an average viab ility of 94.6%. Hepatocytes were then entrapped in type I collagen (3x 10(5) cells/well) and cultured in serum-free media for 5 days. Pig hep atocytes produced stable amounts of albumin and maintained cytochrome P-450 and glucuronidation activity over 5 days, as shown by the metabo lism of lidocaine and 4-methylumbelliferone. These data indicate that pig hepatocytes can be harvested with high yields and can retain viabi lity and differentiated function over at least 5 days of culture, and therefore should prove to be an excellent source of hepatocytes for bi oartificial liver devices.