Duret, C.; Moreno Luqui, Daniel
; Balasiddaiah, A.; Roux, S.; Briolotti, P.; Raulet, E.; Herrero, A.; Ramet, H.; Biron-Andreani, C.; Gerbal-Chaloin, S.; Ramos, J.; Navarro, F.; Hardwigsen, J.; Maurel, P.; Aldabe Arregui, Rafael
; Daujat-Chavanieu, M.
Hepatocyte transplantation is a promising alternative therapy for the treatment
of hepatic failure, hepatocellular deficiency, and genetic metabolic disorders.
Hypothermic preservation of isolated human hepatocytes is potentially a simple
and convenient strategy to provide on-demand hepatocytes in sufficient quantity
and of the quality required for biotherapy. In this study, first we assessed how
cold storage in three clinically safe preservative solutions (UW, HTS-FRS, and
IGL-1) affects the viability and in vitro functionality of human hepatocytes.
Then we evaluated whether such cold-preserved human hepatocytes could engraft and
repopulate damaged livers in a mouse model of liver failure. Human hepatocytes
showed comparable viabilities after cold preservation in the three solutions. The
ability of fresh and cold-stored hepatocytes to attach to a collagen substratum
and to synthesize and secrete albumin, coagulation factor VII, and urea in the
medium after 3 days in culture was also equally preserved. Cold-stored
hepatocytes were then transplanted in the spleen of immunodeficient mice
previously infected with adenoviruses containing a thymidine kinase construct and
treated with a single dose of ganciclovir to induce liver injury. Engraftment and
liver repopulation were monitored over time by measuring the blood level of human
albumin and by assessing the expression of specific human hepatic mRNAs and
proteins in the recipient livers by RT-PCR and immunohistochemistry,
respectively. Our findings show that cold-stored human hepatocytes in IGL-1 and
HTS-FRS preservative solutions can survive, engraft, and proliferate in a damaged
mouse liver. These results demonstrate the usefulness of human hepatocyte
hypothermic preservation for cell transplantation.