Bioengineering livers using organ scaffolds offers an approach to increasing allograft availability for end stage liver disease. Recellularized liver scaffolds will require multiple differentiated cell types to perform necessary physiologic functions. A rodent model of liver bioengineering was developed to study the necessary components for liver recellularization. Biochamber systems were designed and tested to perfuse detergent for decellularization, and subsequent growth medium with oxygenation during recellularization at 37C. 30 experiments were performed to test various cell lines and growth patterns. Biochamber perfusion using 0.1% SDS decellularized livers as confirmed histologically and was most permissive of later cell growth. Peripheral injection of a human hepatocellular cell line (HEPG2) produced cell clusters localized to injection sites. Delivery of HEPG2 cells through portal vein resulted in venous obstruction and failure of diffuse organ proliferation. Bile duct injection of HEPG2 cells demonstrated good proliferative capacity throughout native liver architecture. Primary umbilical vein endothelial cells (HUVEC) administered via portal vein injection created vascular endothelial lining. Simultaneous injection of both cell types via portal and biliary routes resulted in vascular and parenchymal growth patterns. These results support growth of multiple different human-derived cell types in a rodent model, and support continued investigation towards a goal of bioengineering a human liver on a decellularized animal scaffold.

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