APOPTOSIS OF SERUM-FREE C2.8 MOUSE EMBRYO HEPATOCYTIC CELLS CAUSED BY HEPATOCYTE GROWTH-FACTOR DEPRIVATION

C2.8 mouse embryo hepatocytic cells, acutely required exogenous hepatocyte growth factor (HGF) to survive and proliferate in serum-free Dulbecco's modified Eagle's medium supplemented with insulin, transferrin and Na-selenite. Greater than 90% of cultured C2.8 cells died within 48 hours from plating in the absence of HGF. Conversely, HGF prolonged maintenance of life and stimulated cell proliferation. Removal of HGF from the medium of cultures that had grown to confluency, also resulted in a rapid decreased cell survival. In the last circumstance, light microscopic observations revealed, with high frequency, morphological features characteristic of apoptosis. DNA within the affected cells underwent rapid fragmentation, revealed as a ladder of DNA fragments in multiples of about 200 base pairs. HGF prevented loss of cell viability, morphological damages and retarded DNA fragmentation in confluent C2.8 cells. Cycloheximide delayed cell death caused by HGF deprivation.