Kupffer cells are the resident macrophages in the liver and are part of the first line of defense against bacteria and toxins in the gut. Kupffer cells exhibit typical macrophage morphology and can be activated to produce inflammatory cytokines, growth factors and reactive oxygen species. In vivo, prolonged activation of Kupffer cells are associated with many diseases of the liver including Non-alcoholic Fatty Liver Disease (NAFLD), Non-alcoholic Steato Hepatitis (NASH), and liver damage associated with certain bioactivated chemicals. Human Cryopreserved Kupffer cells are isolated and enriched from dissociated human liver tissue and then cryopreserved as passage 0. Characterization of isolated Kupffer cells includes assessing morphology in plated format and evaluating functional responses to LPS stimulation.