Isolation and characterization of parenchymal cells from normal and cirrhotic rat liver

A technique is described for isolation of adult rat hepatocytes from micronodular cirrhotic livers based on a collagenase digestion procedure. Hepatocytes from normal livers and those chronically injured by thioacetamide did not differ with respect to the viability measured by the trypan blue exclusion test or to the cellular concentrations of protein and glycogen, but the triglyceride content of cells from cirrhotic livers was significantly reduced. Hepatocytes isolated from cirrhotic livers are ultrastructurally in a good state of preservation but they appear to be poorer than controls in RER membranes, although the well-preserved mitochondria are somewhat richer in cristae. No differences were detected between the cell preparations in rates of gluconeogenesis and total de novo fatty acid synthesis, but the secretion of newly synthesized fatty acids was significantly reduced in cells from cirrhotic livers. Thus adult rat hepatocytes can be isolated from thioacetamide-induced micronodular cirrhotic livers with high yield and morphological integrity. Differentiated functions are maintained in suspension for at least 4 h.