Hepatocytes from Fibrotic Liver Possess High Growth Potential in Vivo

Abstract

Hepatocyte transplantation is effective for treating liver failure, but healthy donors as a source of hepatocytes are quite limited. The livers of patients with hepatic fibrosis could be an alternative source; however, few reports have examined the nature of hepatocytes from fibrotic livers (f-hepatocytes). In this study, we compared the growth of f-hepatocytes and hepatocytes from normal livers (n-hepatocytes). Hepatocytes were isolated from normal and CCl(4)-treated wild-type Fischer rats that express dipeptidyl dipeptidase IV (DPPIV) gene (DPPIV(+)). The n- and f-hepatocytes proliferated in culture at similar rates. Both types of hepatocytes were transplanted into DPPIV(-) mutant Fischer rats that had been treated with retrorsine to injure the liver and were partially hepatectomized (PHx) before transplantation. Both n- and f-DPPIV(+)-hepatocytes proliferated and formed colonies. The colony sizes of f-hepatocytes 21 days posttransplantation were approximately three times those of n-hepatocytes. The hepatocytes were analyzed using a fluorescence activated cell sorter (FACS). The FACS profile differed between f- and n-hepatocytes: f-hepatocytes were less granular, less autofluorescent, and smaller than n-hepatocytes. These characteristics of f-hepatocytes resembled those reported for small-sized n-hepatocytes (SHs), which are highly proliferative and preferentially express a unique set of 10 SH genes. However, f-hepatocytes preferentially expressed only five of the SH genes. The expression profile of f-hepatocytes was rather similar to that of proliferating n-hepatocytes in the regenerating liver after PHx. The f-hepatocytes were morphologically normal and did not show any preneoplastic phenotype. These normal and proliferative natures of f-hepatocytes in vivo suggest the fibrotic liver as a source of hepatocytes for transplantation.

Construction of a Portal Implantable Functional Tissue-Engineered Liver Using Perfusion-Decellularized Matrix and Hepatocytes in Rats

“…The foremost limiting issue is the identification of a reliable source of large numbers of highly functional liver cells (5), especially various autologous adult stem cell sources such as oval cells, small hepatoblasts, hematopoietic stem cells, and mesenchymal stem cells. We will search for new sources of hepatocytes for transplantation, in particular focusing on the ex vivo genetic modification of patients' own stem or progenitor cells prior to reimplantation (10,16,20,25).…”

Repopulation of the immunosuppressed retrorsine‐treated infant rat liver with human hepatocytes

“…To compare the pro‐proliferative potential of the adult and infant hepatic microenvironment, we calculated the average size of colonies formed by adult rat donor hepatocytes at 3 weeks after transplantation in adult and infant rat models. The average size of a colony is measured in terms of area in the adult RS/PH rat model and the infant RS rat model and was found to be approximately 26 000 μm² and 60 000 μm² (this study), respectively. This result indicates that the pro‐proliferative potential is higher in the infant liver than in its adult counterpart.…”

First approaches for the transplantation of hepatocytes from Wistar rat preneoplastic livers into healthy recipients

“…Group R received 1.5 x 10 5 hepatocytes in 200 µl of Ringer solution. Hepatocyte solutions were injected directly into the splenic pulp using a tuberculin syringe with a G21 (0.8 x 25 mm) needle (18) immediately after partial hepatectomy; 1.5 x 10 5 hepatocytes were used, based on bibliographic references (19) to ensure that a portal thrombosis did not occur. In addition, a simulation of the transplant was performed by injecting the animals (n = 6) with only the vehicle used in the hepatocyte solution (group Sham).…”