Human non-parenchymal liver cells for co-cultivation systems

摘要

Recently, Pfeiffer and colleagues have published protocols that allow the isolation of human hepatocytes and non-parenchymal liver cells from the same donor (Pfeiffer et al., 2014). Cell isolation is performed with resected liver tissue, usually obtained from hepatectomy because of metastasis from colon cancer. Human liver cells are initially isolated by the conventional two-step EDTA/ collagenase perfusion technique. Next, hepatocytes and non-parenchymal cells are separated by low-speed centrifugation. After purification by Percoll density gradient centrifugation, Kupffer cells, sinusoidal endothelial cells and stellate cells are further separated by specific adhesion and by magnetic bead sorting. Typical yields of a single isolation are 1.9x106 Kupffer cells, 2.7x105 sinusoidal endothelial cells and 4.7x105 stellate cells. All cell types can be cultivated either as mono- or co-cultures.