Therapeutic Liver Reconstitution With Murine Cells Isolated Long After Death

Background & Aims

Due to the shortage of donor organs, many patients needing liver transplantation cannot receive one. For some liver diseases, hepatocyte transplantation could be a viable alternative, but donor cells currently are procured from the same sources as whole organs, and thus the supply is severely limited.

Methods

Here, we investigated the possibility of isolating viable hepatocytes for liver cell therapy from the plentiful source of morgue cadavers. To determine the utility of this approach, cells were isolated from the livers of non–heart-beating cadaveric mice long after death and transplanted into fumarylacetoacetate hydrolase–deficient mice, a model for the human metabolic liver disease hereditary tyrosinemia type I and a stringent in vivo model for hepatic cell transplantation.

Results

Surprisingly, complete and therapeutic liver repopulation could be achieved with hepatocytes derived up to 27 hours post mortem.

Conclusions

Competitive repopulation experiments showed that cadaveric liver cells had a repopulation capacity similar to freshly isolated hepatocytes. Importantly, viable hepatocytes also could be isolated from cadaveric primate liver (monkey and human) efficiently. These data provide evidence that non–heart-beating donors could be a suitable source of hepatocytes for much longer time periods than previously thought possible.

Keywords: Fumarylacetoacetate Hydrolase (Fah)-Deficient Mice, Hereditary Tyrosinemia Type I, Liver Disease, Hepatocytes

Abbreviations used in this paper: AAV, adeno-associated virus, Fah, fumarylacetoacetate hydrolase, G6PD, glucose-6-phospho-diesterase, HNF4α, hepatocyte nuclear factor 4-α, NTBC, 2-(2-nitro-4-3 trifluoro-methylbenzoyl)-1,3-cyclohexanedione, PCR, polymerase chain reaction, R26R, Rosa26 reporter mice, Ttr, transthyretin receptor