Small Animal Models of Hepatocyte Transplantation

Abstract

In this chapter, we describe techniques used to determine the efficiency of hepatocyte transplantation in animal models of liver disease. We have included the Gunn rat as a model of an inherited liver disease without hepatocyte damage and Abcb4 knockout mice as a model for an inherited liver disease with hepatocyte damage. Immunodeficient mice are included as an animal model for human hepatocyte transplantation.We describe problems that can be encountered in the maintenance and breeding of Gunn rats and immunodeficient Rag2/gamma common knockout mice. Protocols for the collection of bile in rats and mice are described, and we have also detailed the detection of green fluorescent protein (GFP)-labelled human hepatocytes in immunodeficient mice in this chapter.

Rodent Animal Models for Surrogate Analysis of Cell Therapy in Acute Liver Failure

“…It is assumed that the cells distribute homogeneously in the liver by passage with the blood stream where they are entrapped in the sinusoids and eventually penetrate the endothelia, integrate, proliferate, and spread into the host parenchyma. This concept has been verified in various rodent animal models of ALF (for recent reviews, see Fox and Roy-Chowdhury, 2004; Shafritz et al, 2006; Seppen et al, 2009; Weber et al, 2009). There is one major constraint, which probably seriously hampers the clinical translation of hepatocyte transplantation in ALF.…”

The generation of hepatocytes from mesenchymal stem cells and engraftment into the liver

“…Therefore, rodent animal models aim at providing transplanted cells with a growth advantage over the host hepatocytes. This is achieved by, for example, treatment of the host animals with the pyrrolizidine alkaloid retrorsine, which impairs hepatocyte mitosis, in addition to a proliferative challenge induced by, for example, partial hepatectomy [49][50][51][52][53]. Donor hepatocytes are typically found in the host parenchyma at a rate of 10-15% 2 weeks following transplantation gradually increasing to nearly total replacement after 4-9 months [47,54].…”

Methods of Liver Stem Cell Therapy in Rodents as Models of Human Liver Regeneration in Hepatic Failure

“…18,19 Alu sequence detection by polymerase chain reaction (PCR) and green fluorescent protein (GFP) fusions are other useful techniques for tracking and locating transplanted cells. 9,19,21,28 Functional assay of transplanted cells Characterization of cell viability and cell identity after administering the cells is very important in the long-term follow-up study of liver cell therapy. Mortality rate is a reliable indicator of transplant functionality and overall recipient health.…”

“…The major candidate cells for cell therapy are primary autologous mesenchymal cells, genetically identical or syngeneic cells, immortalized hepatocytes 7 and stem cells. 2,8 Currently, cellular transplantation is developing for liver damage therapy 9 and in comparison to liver transplantation has many advantages including overcoming the shortage of liver donors 1,6,10 and immunological rejection, 1,10 and a minimally invasive approach. 7 Research in the area of cell replacement therapy in the liver, particularly with stem cells, is at the beginning stages.…”