Culturing hepatocytes on different extracellular matrix (ECM) substrata including tissue culture plastic, type I collagen, Engelbreth-Holm-Swarm (EHS) gel and poly-N-p-vinylbenzyl-D-lactonamide (PVLA) regulated levels of mRNAs for cytoskeleton and liver-specific genes. In hepatocytes on EHS gel, the ratio of albumin/beta-actin in mRNA levels was high and serially increased during the culture period, while the ratio was low and declined in cells on plastic substratum, collagen or PVLA. The changes in cellular levels of albumin mRNA which were regulated by ECM corresponded with those in two liver-specific transcription factors, hepatocyte nuclear factors-1 and -4, which control the transcription of liver-specific genes. These results suggest that cell-matrix interaction may determine and maintain the differentiated phenotype of hepatocytes by regulating liver-specific transcription factors.