Portal venous repopulation of decellularised rat liver scaffolds with syngeneic bone marrow stem cells.

Harper, Simon; Hoff, Mekhola; Skepper, Jeremy; Davies, Susan; Huguet, Emmanuel

Liver transplantation is the only life-saving treatment for end stage liver failure but is limited by the organ shortage and consequences of immunosuppression. Repopulation of decellularized scaffolds with recipient cells provides a theoretical solution, allowing reliable and timely organ sourcing without the need for immunosuppression. Recellularisation of the vasculature of decellularised liver scaffolds was investigated as an essential pre-requisite to the survival of other parenchymal components. Liver decellularisation was carried out by portal vein perfusion using a detergent-based solution. Decellularised scaffolds were placed in a sterile perfusion apparatus consisting of a sealed organ chamber, functioning at 37°C in normal atmospheric conditions. The scaffold was perfused via portal vein with culture medium. A total of 107 primary cultured bone marrow stem-cells, selected by plastic adherence, were infused into the scaffold, after which repopulated scaffolds were perfused for up to 30 days. The cultured stem cells were assessed for key marker expression using FACS and recellularised scaffolds were analysed by light, electron and immunofluorescence microscopy. Stem-cells engrafted in portal, sinusoidal and hepatic vein compartments, with cell alignment reminiscent of endothelium. Cell-surface marker expression altered following engraftment, from haematopoietic to endothelial phenotype and engrafted cells expressed sinusoidal endothelial endocytic receptors (mannose, Fc, stabilin receptors). These results represent one step towards complete recellularisation of the liver vasculature and progress towards the objective of generating transplantable neo-organs.