Human Hepatocytes Can Repopulate Mouse Liver: Histopathology of the Liver in Human Hepatocyte-Transplanted Chimeric Mice and Toxicologic Responses to Acetaminophen

Although multiple determinants for hepatitis C virus (HCV) infection are known, it remains partly unclear what determines the human specificity of HCV infection. Presumably, the presence of appropriate entry receptors is essential, and this may explain why HCV is unable to infect nonhuman hepatocytes. However, using mice with chimeric human livers, we show in this study that the presence of human hepatocytes, and therefore human entry receptors, is not sufficient for HCV infection. In successfully transplanted SCID/Alb-uPA mice, infection with HCV is reliable only when ∼70–80% of the liver consists of human hepatocytes. We show that chimeric mice, which are hard to infect with HCV, have significant groups of human hepatocytes that are readily infected with hepatitis B virus. Thus it is unlikely that the lack of infection with HCV can simply be attributed to low hepatocyte numbers. We investigated whether the humanization of lipoprotein profiles is positively associated with infection success. We show that the lipoprotein profiles of chimeric mice become more human-like at high levels of engraftment of human hepatocytes. This and expression of markers of human lipoprotein biosynthesis, human apolipoprotein B (ApoB) and cholesterol ester transfer protein (CETP), show a strong positive correlation with successful infection. Association of HCV in the blood of chimeric mice to ApoB-containing lipoproteins is comparable to association of HCV in patient serum and provides further support for a critical role for ApoB-containing lipoproteins in the infectious cycle of HCV. Our data suggest that the weakest link in the HCV infection chain does not appear to be the presence of human hepatocytes per se. We believe that HCV infection also depends on the presence of sufficient levels of human lipoproteins.

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Characterization and applications of chimeric mice with humanized livers for preclinical drug development

Laboratory Animal Research ◽

10.1186/s42826-019-0032-y ◽

2020 ◽

Vol 36 (1) ◽

Cited By ~ 4

Author(s):

Chise Tateno ◽

Yuha Kojima

Keyword(s):

Liver Function ◽

Drug Development ◽

Mass Production ◽

Human Hepatocytes ◽

In Vitro Studies ◽

New Drugs ◽

Pharmaceutical Companies ◽

Chimeric Mice ◽

Preclinical Drug Development

AbstractWe have succeeded in stable mass production of chimeric PXB-mice, whose liver is repopulated by human hepatocytes at a ratio of more than 70%, and we are providing these mice to academia and pharmaceutical companies to support the development of new drugs or studies of liver function. Furthermore, we isolated human hepatocytes, called PXB-cells, from the chimeric mice, and provide them for clients weekly for in vitro studies. In this review, we summarize the existing characterizations of PXB-mice and PXB-cells and their present and future applications.

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Synthesis of Factor VIII in Human Hepatocytes in Culture

Thrombosis and Haemostasis ◽

10.1055/s-0038-1646977 ◽

1988 ◽

Vol 60 (03) ◽

pp. 387-391 ◽

Cited By ~ 8

Author(s):

J Ingerslev ◽

B Sloth Christiansen ◽

L Heickendorff ◽

C Munck Petersen

Keyword(s):

Factor Viii ◽

Light Chain ◽

Gradient Centrifugation ◽

Human Fibroblasts ◽

Human Hepatocytes ◽

Human Hepatocyte ◽

Blood Monocytes ◽

Hep G2 ◽

Human Hepatoma Cells ◽

Sds Page

SummaryAlthough several investigators have attempted to identify the site of synthesis of factor VIII (FVIII), the cellular species responsible for maintenance of plasma FVIII has not been clearly defined. Indications point at hepatocytes and certain endothelial cells. The present study investigated the FVIII coagulant antigen (VIII : Ag) of hepatocytes obtained by two-step collagenase digests of human liver pieces. Following Percoll gradient centrifugation, less than 1% of cells harvested were non-parenchymal. Lysates of freshly isolated and purified hepatocytes contained 165–250 mU of VIII: Ag/106 cells as defined by a two-site ELISA employing a haemophilic antibody against human FVIII. This material contained a single peak of VIII: Ag polypeptides as jugded from the VIII: Ag ELISA profile of Mono-Q fast protein liquid chromatography fractions. A haemophilic antibody specific for epitopes of the light chain of FVIII, employed in immunoisolation of VIII : Ag in lysate of human hepatocytes, extracted a polypeptide pattern that was studied in a reduced SDS-PAGE electrophoresis gel and compared to that of immunoisolate from normal plasma. After electroblotting onto nitrocellulose and reaction with a monoclonal antibody towards the light chain of FVIII, the appearance of a doublet at 78–79 kDa in both these materials indicated the presence of the light chain of FVIII in human hepatocyte lysate. During culture, human hepatocytes secreted 20–80 mU of VIII: Ag per 1 × 106 cells per 24 hours. Further, a significant secretion of VIII: Ag was found in media of cultured human hepatoma cells, Hep-G2, whereas human blood monocytes and human fibroblasts did not secrete detectable VIII: Ag. In all of these cell cultures, vWf : Ag was indetectable or present as trace. Our results suggest that the human hepatocyte is a production site of FVIII.

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Assessing the compatibility of primary human hepatocyte culture within porous silk sponges

RSC Advances ◽

10.1039/d0ra04954a ◽

2020 ◽

Vol 10 (62) ◽

pp. 37662-37674

Author(s):

David A. Kukla ◽

Whitney L. Stoppel ◽

David L. Kaplan ◽

Salman R. Khetani

Keyword(s):

Extracellular Matrix ◽

Extracellular Matrix Proteins ◽

Human Hepatocytes ◽

Human Hepatocyte ◽

Matrix Proteins ◽

Hepatocyte Culture ◽

Primary Human Hepatocytes ◽

Primary Human Hepatocyte ◽

Silk Scaffolds ◽

Parenchymal Cells

Porous silk scaffolds hybridized with extracellular matrix proteins are useful for culture of primary human hepatocytes ± supportive non-parenchymal cells.

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Comparison of the Hepatic Effects of Phenobarbital in Chimeric Mice Containing Either Rat or Human Hepatocytes With Humanized Constitutive Androstane Receptor and Pregnane X Receptor Mice

Toxicological Sciences ◽

10.1093/toxsci/kfaa125 ◽

2020 ◽

Vol 177 (2) ◽

pp. 362-376

Author(s):

Tomoya Yamada ◽

Ayako Ohara ◽

Naoya Ozawa ◽

Keiko Maeda ◽

Miwa Kondo ◽

...

Keyword(s):

Gene Expression ◽

Liver Tumor ◽

Constitutive Androstane Receptor ◽

Pregnane X Receptor ◽

Rat Hepatocytes ◽

Global Gene Expression ◽

Human Hepatocytes ◽

Chimeric Mouse ◽

Chimeric Mice ◽

Wild Type

Abstract Using a chimeric mouse humanized liver model, we provided evidence that human hepatocytes are refractory to the mitogenic effects of rodent constitutive androstane receptor (CAR) activators. To evaluate the functional reliability of this model, the present study examined mitogenic responses to phenobarbital (PB) in chimeric mice transplanted with rat hepatocytes, because rats are responsive to CAR activators. Treatment with 1000 ppm PB for 7 days significantly increased replicative DNA synthesis (RDS) in rat hepatocytes of the chimeric mice, demonstrating that the transplanted hepatocyte model is functionally reliable for cell proliferation analysis. Treatment of humanized CAR and pregnane X receptor (PXR) mice (hCAR/hPXR mice) with 1000 ppm PB for 7 days significantly increased hepatocyte RDS together with increases in several mitogenic genes. Global gene expression analysis was performed with liver samples from this and from previous studies focusing on PB-induced Wnt/β-catenin signaling and showed that altered genes in hCAR/hPXR mice clustered most closely with liver tumor samples from a diethylnitrosamine/PB initiation/promotion study than with wild-type mice. However, different gene clusters were observed for chimeric mice with human hepatocytes for Wnt/β-catenin signaling when compared with those of hCAR/hPXR mice, wild-type mice, and liver tumor samples. The results of this study demonstrate clear differences in the effects of PB on hepatocyte RDS and global gene expression between human hepatocytes of chimeric mice and hCAR/hPXR mice, suggesting that the chimeric mouse model is relevant to humans for studies on the hepatic effects of rodent CAR activators whereas the hCAR/hPXR mouse is not.

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