Sinusoidal cells were isolated by collagenase perfusion and metrizamide gradient centrifugation, from liver resected for partial hepatectomy performed under warm ischemic conditions. Kupffer cells were then separated from this population by centrifugal elutriation. Isolated Kupffer cells showed good viability, with the typical features of Kupffer cells and were engaged in the endocytosis of foreign particles. They showed numerous morphological criteria of activation. However, cultured Kupffer cells were no longer in an activated state. Kupffer cells were preserved in maintenance cultures for 2 weeks. Purity of these cultures was to 93-97%. During culture, Kupffer cells retained their ultrastructural characteristics and were active in the endocytosis of latex beads and opsonized zymosan particles. It is thus possible that partial hepatectomy performed under warm ischemia could provide valuable material for the study of Kupffer cells in vitro.