Assessing Rat Liver-Derived Biomatrix for Hepatic Tissue Engineering with Human Fetal Liver Stem Cells

Decellularized organs with preserved architecture and vasculature have been created to produce customized bioengineered organs Identification of appropriate xenogeneic scaffolds which can support attachment proliferation and differentiation of human cells is important for studying human organ regeneration Here we produced D decellularized scaffolds from rat n livers preserving the native morphology and vascular structures with complete removal of cellular and nuclear materials The ability of human liver sinusoidal endothelial cells LSEC and two previously well characterized human fetal liver progenitor cell HFLPC lines SV and hFL TERT to repopulate the acellular rodent liver tissues was evaluated We found that although the major extracellular proteins were preserved cytokines growth factors tested were significantly decreased in the decellularized livers The portal veins were fragile at the end of the decellularization process resulting in technical problems during recellularization Even though the vascular network looked preserved after decellularization leakage was observed in majority of the livers Only in four of the recellularized rat scaffolds attachment of human HFLPC was seen However the cells were not evenly distributed in the scaffolds but found scattered as colonies in the rat parenchyma and blood vessels Approximately of the human cells were viable at the end of recellularization with stable expression of human mitochondria The cells also showed expression of the endothelial marker L SIGN and weak expression of human albumin Few cells expressed hepatocyte specific markers cytokeratin CK and CK but with high variablilty while but no expression of the biliary cell marker CK was found A few cells expressed CYP A but CYP A was not found Based on our experience and results we do not believe that decellularized rat liver scaffolds are the ideal scaffolds for reproducible recellularization with human liver stem cells required for studying human liver regeneration or as in vitro models for pharmaceutical toxicity