Analysis of the functional integrity of cryopreserved human liver cells including xenografting in immunodeficient mice to address suitability for clinical applications.
BACKGROUND
The availability of well-characterized human liver cell populations that can be frozen and thawed will be critical for cell therapy. We addressed whether human hepatocytes can recover after cryopreservation and engraft in immunodeficient mice.


METHODS
We isolated cells from discarded human livers and studied the properties of cryopreserved cells. The viability of thawed cells was established with multiple in vitro assays, including analysis of liver gene expression, ureagenesis, cytochrome P450 activity, and growth factor-induced cell proliferation. The fate of transplanted cells was analysed in immunodeficient NOD-SCID mice.


RESULTS
After thawing, the viability of human hepatocytes exceeded 60%. Cells attached to culture dishes, proliferated following growth factor stimulation and exhibited liver-specific functions. After transplantation in NOD-SCID mice, cells engrafted in the peritoneal cavity, a heterologous site, as well as the liver itself, retained hepatic function and proliferated in response to liver injury. Transplanted hepatocytes were integrated in the liver parenchyma. Occasionally, transplanted cells were integrated in bile ducts.


CONCLUSIONS
Cryopreserved human liver cell showed the ability to retain functional integrity and to reconstitute both hepatic and biliary lineages in mice. These studies offer suitable paradigms aimed at characterizing liver cells prior to transplantation in people.
Cho
Jae-Jin
JJ
Department of Medicine, Marion Bessin Liver Research Center, Albert Einstein College of Medicine, Bronx, NY 10461, USA.
Joseph
Brigid
B
Sappal
Baljit Singh
BS
Giri
Ranjit K
RK
Wang
Richard
R
Ludlow
John W
JW
Furth
Mark E
ME
Susick
Robert
R
Gupta
Sanjeev
S
D000818:Animals; D001681:Biological Assay / Q000379:methods; D002455:Cell Division; D002465:Cell Movement; D002470:Cell Survival; D015925:Cryopreservation; D006085:Graft Survival; D022781:Hepatocytes / Q000166:cytology* / Q000637:transplantation*; D006801:Humans; D008099:Liver / Q000166:cytology; D051379:Mice; D016688:Mice, Inbred NOD; D016513:Mice, SCID; D014183:Transplantation, Heterologous