ABSTRACT
This chapter demonstrates that valuable insight can be gained regarding cellular dedifferentiation and effects of compounds on hepatocytes via gene expression analysis. It provides a comprehensive, temporal gene-expression-based analysis of a traditional monolayer cultured hepatocyte system. The use of cultured primary hepatocytes to model hepatotoxicity has proven to be a valuable tool; however, questions remain with regard to functional differences observed in primary hepatocytes relative to the intact liver. Parenchymal hepatocytes are complex, highly differentiated cells that comprise over 80% of the normal liver mass and 60% of the total liver cell population. Hepatocytes were isolated from 8- to 10-week-old male rats using a modified two-stage portal vein perfusion technique. Whole liver samples were collected prior to cannulation by ligating and removing the caudate lobe. Isolated hepatocytes were collected for analysis at various time points throughout the isolation process. Several approaches were used to verify the array data collected in the hepatocyte differentiation study.
