**10. Conclusion**

Mito et al. (1978) completed the first successful hepatocyte transplantation in the rat spleen 33 years ago. Since then, cosiderable progress has been made in the field of hepatic cell transplantation research. Notably, it has been demonstrated that hepatocytes can be transplanted into the liver and can regenerate the diseased host liver. The use of hepatocyte transplantation is therefore expected to expand as a therapy for human liver disease. For cell transplantation to be used as a human therapy, further investigation is required to address number of problems including efficient ways to suppress the innate and acquired immune responses to the transplanted cells and the short- and long-term risks such as malignant transformation. The F344/F344-alb model provides a useful tool for studies of hepatocyte transplantation in the liver because it can be used to accurately trace the fate and functionality of the transplanted cells. Bone marrow cell transplantation may facilitate cell transplantation therapy via the hepatogenic potential of hematopoietic cells, the putative protective paracrine actions in the liver and the induction of donorspecific immuno-tolerance. A shift from orthotropic liver transplantation, if in part, to hepatocyte transplantation will yield new opportunities to develop therapies for human liver disease.

### **11. Acknowledgements**

This work was supported by the grants from the Japanese Ministry of Science, Education, Sports and Culture.

### **12. References**

Arikura, J., Inagaki, M., Huiling, X., Ozaki, A., Onodera, K., Ogawa, K. & Kasai, S. (2004). Colonization of albumin-producing hepatocytes derived from transplanted F344 rat bone marrow cells in the liver of congenic Nagase's analbuminemic rats. *Journal of Hepatology*, Vol. 41, No. 2, pp. 215-221.

levels. Although slight inflammatory cell infiltration was observed in the portal areas of the recipient livers, no inflammatory changes were detected in the areas repopulated with albumin-positive hepatocytes. When quantitative PCR was used to test bone marrow reconstitution in the recipient F344-alb rats for the normal and analbuminemic albumin gene sequences, 6 of 6 F344-alb rats showed bone marrow reconstitution after intra-bone marrow injection (Figure 6C). In contrast, only 1 of 6 F344-alb rats that underwent intravenous bone marrow cell transfusion showed bone marrow reconstitution. The albumin-positive hepatocyte repopulation and the increase in serum albumin levels depended completely on

These results indicate that liver repopulation via allogeneic hepatocyte transplantation without the use of immunosuppressants is possible if the recipient bone marrow is efficiently reconstituted using donor bone marrow cells. Intra-bone marrow injection of bone marrow cells induces bone marrow reconstruction in a manner that is more efficient

Mito et al. (1978) completed the first successful hepatocyte transplantation in the rat spleen 33 years ago. Since then, cosiderable progress has been made in the field of hepatic cell transplantation research. Notably, it has been demonstrated that hepatocytes can be transplanted into the liver and can regenerate the diseased host liver. The use of hepatocyte transplantation is therefore expected to expand as a therapy for human liver disease. For cell transplantation to be used as a human therapy, further investigation is required to address number of problems including efficient ways to suppress the innate and acquired immune responses to the transplanted cells and the short- and long-term risks such as malignant transformation. The F344/F344-alb model provides a useful tool for studies of hepatocyte transplantation in the liver because it can be used to accurately trace the fate and functionality of the transplanted cells. Bone marrow cell transplantation may facilitate cell transplantation therapy via the hepatogenic potential of hematopoietic cells, the putative protective paracrine actions in the liver and the induction of donorspecific immuno-tolerance. A shift from orthotropic liver transplantation, if in part, to hepatocyte transplantation will yield new opportunities to develop therapies for human

This work was supported by the grants from the Japanese Ministry of Science, Education,

Arikura, J., Inagaki, M., Huiling, X., Ozaki, A., Onodera, K., Ogawa, K. & Kasai, S. (2004).

Colonization of albumin-producing hepatocytes derived from transplanted F344 rat bone marrow cells in the liver of congenic Nagase's analbuminemic rats. *Journal of* 

bone marrow reconstitution by the donor bone marrow cells.

than does intravenous transfusion.

**10. Conclusion** 

liver disease.

**11. Acknowledgements** 

*Hepatology*, Vol. 41, No. 2, pp. 215-221.

Sports and Culture.

**12. References** 


Analbuminemic Rat Model for Hepatocyte Transplantation 139

Ohta, T., Ogawa, K. & Nagase, S. (1993a). Increase in albumin mRNA by repeated

Ohta, T., Ogawa, K. & Nagase, S. (1993b). Elevation of serum albumin by intrahepatic

Ohta, T., Ogawa, K. & Nagase, S. (1994). Analbuminemia does not significantly influence

Rothschild, M.A., Oratz, M., & Schreiber, S.S. (1988). Serum albumin. *Hepatology,* Vol. 8, No.

Sato, K. (1989). Glutathione transferases as markers of preneoplasia and neoplasia*. Advance* 

Serandour A.L., Loyer, P., Garnier, D., Courselaud, B., Théret, N., Glaise., D, Guguen-

Shalaby, F. & Shafritz, D.A. (1990). Exon skipping during splicing of albumin mRNA

Takahashi, M., Shumiya, S., Maekwa A., Hayashi, Y. & Nagase, S. (1988). High susceptibility

Theise, N.D., Nimmakayalu, M., Gardner, R., PB. Illei, P.B., Morgan, G., Tepeman, L.,

Vidal, I., Blanchard, N., Alexandre, E., Gandillet, A., Chenard-Neu, M.P., Staedtler, F.,

treatment. *Cell Transplanataion,* Vol. 17, No. 5, pp. 507-524.

Guillouzo, C. & Corlu, A. (2005). TNF-mediated extracellular matrix remodeling is required for multiple division cycles in rat hepatocytes. *Hepatology,* Vol. 41, No. 3,

precursors in Nagase analbuminemic rats. *Proceeding of National Academy of Science* 

of analbuminemic congenic strain of rats with an F344 genetic background to induced bladder cancer and its possible mechanism. *Japanese Journal of Cancer* 

Heneqariu, O. & Krause, D.S. (2000). Liver from bone marrow in humans.

Schumacher, M., Bachellier,P., Jaeck, D., Firat, H., Heyd, B. & Richer, L. (2008). Improved xenogenic hepatocyte implantation into nude mouse liver parenchyma with acute liver failure when followed by repeated anti-Fas antibody (Jo2)

*Biophysical Research Communications,* Vol. 197, No. 3, pp. 1103-1110.

analbuminemic mutation. *Carcinogenesis,* Vol. 15, No. 2, pp. 227-231. Okazaki, S., Hisha, H., Mizokami, T., Takaki, T., Wang, T., Song, C., Li, Q., Kato, J.,

Roy-Chowdhury, N. & Roy-Chowdhury, J. Hepatocyte transplantation.

*Gastroenterology,* Vol. 132, No. 3, pp. 1077-1087.

No. 2, pp. 601-609.

17, No. 4, pp. 629-640.

*http://www.uptodate.com/contents/*

*in Cancer Research,* Vol. 52, pp. 205-255.

*USA,* Vol. 87, No. 4, pp. 2652-2656.

*Research,* Vol. 79, No. 6, pp. 705-709.

*Hepatology*, Vol. 32, No. 1, pp. 11-16, 2000.

2, pp. 385-401.

pp.478-486.

acetylaminofluorene/partial hepatectomy-induced liver regeneration.

intrahepatic transplantation of F344 rat hepatocytes into the liver of congenic analbuminemic rats. *Biochemical and Biophysical Research Communications,* Vol. 194,

transplantation of albumin-producing cells does not correct quantitative abnormalities of non-albumin proteins in analbuminemic rats. *Biochemical and* 

hepatocarcinogenesis on comparing F344 rats and a congenic line carrying the

Kamiyama, Y. & Ikehara, S. (2008). Successful acceptance of adult liver allografts by intra-bone marrow-bone marrow transplantation. *Stem Cells and Development*, Vol.


Inagaki, M., Furukawa, H., Satake, Y., Okada, Y., Chiba, S., Nishikawa, Y. & Ogawa, K.

Inoue, M. (1985) Metabolism and transport of amphipathic molecules in analbuminemic ras

Kakizoe, T., Komatsu, H., Honma, Y., Niijima, Y., Kawachi, T., Sugimura, T. & Nagase, S.

Kaneko, T., Shima, H., Esumi, H., Ochiai, M., Nagase, S., Sugimura, T. & Nagao, M. (1991).

Kim, S.H., Kim, J.H. & Akaike, T. (2003). Regulation of cell adhesion signaling by synthetic

Laconi, E., Oren, R., Mukhopadyay, D.K., Hurston, E., Laconi, S., Pani, P., Dabeva, M.D. &

Makino, R., Sato, S., Esumi, H., Negishi, C., Takano, M., Sugimura, T., Nagase, S. & Tanaka,

Mao, X., Fujikawa, Y. & Orikin, S.H. (1999). Improved reporter strain for monitoring Cre

Merion, R.M. (2010). Current status and future of liver transplantation. *Seminar of Liver* 

Mito, M., Ebata, H., Kusano, M., Onishi, T., Saito, T. & Sakamoto, S. (1978). Morphology and

Nagase, S., Shimamune, K. & Shumiya, S. (1979). Albumin-deficient rat mutant. *Science,* Vol.

Nishikawa, S., Ohta, T., Ogawa, K. & Nagase, S. (1994). Reversion of altered phenotype in

Ogawa, K., Ohta, T., Inagaki, M. & Nagase, S. (1993). Identification of F344 rat hepatocytes

Oh, S.H., Witek, R.P., Bae, S.H., Zheng, D., Jung, Y., Piscaqlia, A.C. & Petersen, B.E. (2007).

transplantation. *Laboratory Investigation*, Vol. 70, No. 6, pp. 925-932.

chain reaction. *Transplantation,* Vol. 56, No.1, pp. 9-15.

*Transplantation,* Vol. 20, No. 9, pp.1479-1489.

*Journal of Cancer,* Vol. 45, No. 3, pp. 474-476.

pp. 433-439.

Vol. 153, No. 1, pp. 319-329.

*Research,* Vol. 77, No. 2, pp. 153-159.

*Disease,* Vol. 30, No. 4, pp. 411-421.

Vol. 28, No. 6, pp. 499-505.

205, No. 4406, pp. 590-591.

*Science USA,* Vol. 96, No. 9, pp. 5037-5042.

and human subjects. *Hepatology*, Vol. 5, No. 5, pp.892-898.

*Academy of Science USA,* Vol. 88, No. 4, pp. 2807-2811.

(2011). Replacement of liver parenchyma in analbuminemic rats with allogenic hepatocytes is facilitated by intrabone marrow-bone marrow transplantation. *Cell* 

(1982). High susceptibility of analbuminemic rats to induced bladder cancer. *British* 

Marked increases of two kinds of two-exon-skipped albumin mRNAs with aging and their further increase by treatment with 3'-methyl-4 dimethylaminoazobenzene in Nagase analbuminemic rats. *Proceeding of National* 

glycopolymer matrix in primary cultured hepatocyte. *FEBS Letters,* Vol. 553, No. 3,

Shafritz, D.A. (1998). Long-term, near total liver replacement by transplantation of isolated hepatocytes in rats treated with retrorsine. *American Journal of Pathology*,

H. (1986). Presence of albumin-positive cells in the liver of analbuminemic rats and their increase on treatment with hepatocarcinogens. *Japanese Journal of Cancer* 

recombinase-mediated DNA excisions in mice. *Proceeding of National Academy of* 

function of isolated hepatocytes transplanted into the rat spleen. *Transplantation,*

primary cultured rat hepatocytes after intrahepatic and intrasplenic

transplanted within the liver of congenic analbuminemic rats by the polymerase

Bone marrow-derived hepatic oval cells differentiate into hepatocytes in 2-

acetylaminofluorene/partial hepatectomy-induced liver regeneration. *Gastroenterology,* Vol. 132, No. 3, pp. 1077-1087.


**8** 

*CHU Necker, France* 

**Rodent Models with Humanized Liver:** 

*INSERM, U845, Pathogenèse des Hépatites Virales B et Immunothérapie,* 

*Université Paris Descartes, Sorbonne Paris Cité, Faculté de Médecine René Descartes,* 

The recent development of small animal models for experimental hepatotropic infection has opened new perspectives for the evaluation of novel therapeutic and/or prophylactic compounds against hepatitis B virus (HBV), hepatitis C virus (HCV) and Plasmodium falciparum, three major hepatic pathogens responsible for millions of deaths each year. Indeed, till now in vitro and in vivo models have their limitations. As example, primary human hepatocytes (PHH) are susceptible to infection by HBV (Gripon et al 1988), HCV (Fournier et al 1998) and by sporozoites (the hepatic stage of Plasmodium falciparum) (Mazier et al 1985), but are hampered by a rapid dedifferentiation of the PHH (the loss of differentiation leads to a loss of susceptibility to infection) and the difficulties of obtaining fresh cells. In vivo, the chimpanzee constitutes the best non-human primate which can be used for studies of HBV, HCV and Plasmodium falciparum (Dandri et al 2005b; Kremsdorf & Brezillon 2007; Moreno et al 2007), but multiple drawbacks, including ethical issues, the inability to produce numerous progeny in a short time (long gestation periods) and

For a long time, liver cell transplantation was just a dream; fortunately, experimental biology as led researchers to create new challenging mouse models. Indeed, generation of new mouse models for human hepatocyte transplantation have permitted, for the first time, experimental manipulations of human hepatotropic pathogens of man which are immediate problems of human health, as well as the study of cell transplantation in a regenerative medicine perspective. Here, we will focus on the development of humanized mice models

Few papers laid the foundations for the entire field of liver cell transplantation in mouse. They described and applied a genetic-based animal model for competitive liver regeneration where exogenous transplanted hepatocytes have a selective advantage and can replace the diseased tissue. Two mice models were described: transgenic mice expressing high levels of uPA (urokinase-Plasminogen Activator) (Rhim et al 1994) and mice deficient for the fumaryl

exorbitant housing and breeding costs render difficult the accessibility.

using hepatocyte transplantation to study the three major hepatic pathogens.

**2. Transplanted hepatic cells can replace a diseased liver in mice** 

acetoacetate hydrolase (FAH) (Grompe et al 1993) (Fig. 1).

**1. Introduction** 

**A Tool to Study Human Pathogens** 

Ivan Quétier, Nicolas Brezillon and Dina Kremsdorf

