University Medicine Berlin, Department for General, Visceral and Transplantation Surgery, led by Dr. Georg Damm (and Prof. Dr. Daniel Seehofer) have established a protocol for an uncomplicated isolation of primary human hepatocytes (PHH), Kupffer cells (KC), liver endothelial cells (LEC), and human Stellate cells (HSC) from human donor tissue. Liver cells were isolated from the tissue using a two-step EDTA/collagenase perfusion technique, followed by a separation of PHH and different non-parenchymal cell (NPC) fractions through Percoll density gradient centrifugation, an adherence separation step (KC) and magnetic activated cell sorting (HSC, LEC). All isolated cell fractions were identified by specific morphological and functional characteristics and were examined during a defined cultivation time.
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