Hepatocyte transplantation program: Lessons learned and future strategies

Figure 1 Scheme of human hepatocyte isolation from unused donor liver.

Figure 2 Liver from a neonatal donor to be used for hepatocyte isolation.

Figure 3 Optimization of the process of hepatocyte isolation. When livers weighing 1.5-3.5 kg are available, a better recovery was achieved by dividing the hepatic parenchyma (0.5-1 kg) and perfusing each lobe.

Figure 4 Split liver sutured the arterial, portal and suprahepatic branches in the cut surface. A: The application TachoSil^®, a hemostatic sealant, avoids leakage of the collagenase; B: Cannulation of the hepatic veins allows using bigger calibre cannulas increasing the flow of the perfusates.

Figure 5 Hepatocyte isolation and transplantation process. A: Perfusion of the liver at 37 °C; B: Liver after digestion by collagenase perfusion; C: Purification of hepatocytes by low-speed centrifugation; D and E: Hepatocyte suspension is washed twice and pelleted by low-speed centrifugation; F: Freshly isolated hepatocyte suspension; G: Cell thawing by immersing the bags in a fixed-temperature bath at 37 °C; H: Hepatocyte suspension in infusion medium ready to be transplanted; I: Infusion of hepatocytes to the receptor patient.