Decellularized Liver Matrix as a Carrier for Transplantation of Human Fetal and Primary Hepatocytes in Mice

摘要

Transplantation of primary hepatocytes has been shown to augment the function of damaged liver and to “bridge” patients to liver transplantation. However, primary hepatocytes often have low levels of engraftment and short survival after transplantation. To explore the potential benefits of using decellularized liver extracellular matrix (DLM) as a carrier for hepatocyte transplantation, DLM from the whole mouse liver was generated. Immortalized human fetal hepatocytes (FH-hTERT) or primary human hepatocytes were infused into DLM, which was then implanted into the omentum of immuno-deficient NOD/SCID/IL2rγ−/− or NOD/SCID/MPS VII mice. The removal of endogenous cellular components and the preservation of the extracellular matrix proteins and vasculature were demonstrated in the resulting DLM. Bioluminescent imaging revealed that FH-hTERT transduced with a lentiviral vector expressing firefly luciferase survived in the DLM for 8 weeks after peritoneal implantation; whereas, the luciferase signal from FH-TERT rapidly declined in control mice 3–4 weeks after transplantation via splenic injection or with omental implantation after Matrigel encapsulation. Furthermore, primary human hepatocytes reconstituted in the DLM not only survived 6 weeks after transplantation, but also maintained their function, as demonstrated by mRNA levels of albumin and cytochrome P450 subtypes (CYP3A4, CYP2C9 and CYP1A1) similar to freshly isolated human primary hepatocytes. In contrast, when human primary hepatocytes were transplanted into mice via splenic injection, they failed to express CYP3A4, although they expressed albumin. In conclusion, decellularized liver extracellular matrix provides an excellent environment for long-term survival and maintenance of hepatocyte phenotype after transplantation.