Construct Of Bioartificial Liver And Perioperative Application Of Artificial Liver Support System In Orthotopic Liver Transplantation

PART I : Large-scale isolation of primary human hepatocytesPurpose: To develop a suitable method for large scale isolation of primary human hepatocytes.and to investigate the morphologic features and metabolic activity of isolated mass of primary human hepatocytes in vitro culture system. Methods: Human hepatocytes were isolated by two-step extracorporeal perfusion with ethylene diamineteraacetic acid and collagenase solutions. The harvested hepatocytes were inoculated in the fresh William's E medium supplemented with lOOmL/L fetal calf serum and so on. The morphologic features of cultured hepatocytes were recorded. The biosynthesis function and cellular damage of hepatocytes during different cultural intervals were also evaluated. Results: The harvested primary hepatocytes consisted of more than 95% of the total isolated mass liver cells with the cellular viability as high as 94% to 98%, and gave an average yield of 2.5 to 5.8X107 cells per gram. Primary hepatocytes in vitro cultured possessed the features of morphology of normal human primary hepatocytes and have been identified to have the active metabolic and synthetic functions during the initial 14 days. Conclusions: This method allows the high yield harvest and steady proliferation of sufficient mass primary human hepatocytes with high viability and normal metabolic activity in vitro culture system, which may be served as the better supporting model of bioartifical liver devices. Key words: hepatocytes;cell isolation;cell;cultured...