Morphological and microarray analyses of human hepatocytes from xenogeneic host livers

Abstract

We previously produced mice with human hepatocyte (h-hep) chimeric livers by transplanting h-heps into albumin enhancer/promoter-driven urokinase-type plasminogen activator-transgenic severe combined immunodeficient (SCID) mice with liver disease. The chimeric livers were constructed with h-heps, mouse hepatocytes, and mouse hepatic sinusoidal cells (m-HSCs). Here, we investigated the morphological features of the chimeric livers and the h-hep gene expression profiles in the xenogeneic animal body. To do so, we performed immunohistochemistry, morphometric analyses, and electron microscopic observations on chimeric mouse livers, and used microarray analyses to compare gene expression patterns in hepatocytes derived from chimeric mouse hepatocytes (c-heps) and h-heps. Morphometric analysis revealed that the ratio of hepatocytes to m-HSCs in the chimeric mouse livers were twofold higher than those in the SCID mouse livers, corresponding to twin-cell plates in the chimeric mouse liver. The h-heps in the chimeric mouse did not show hypoxia even in the twin-cell plate structure, probably because of low oxygen consumption by the h-heps relative to the mouse hepatocytes (m-heps). Immunohistochemical and electron microscopic examinations revealed that the sinusoids in the chimeric mouse livers were normally constructed with h-heps and m-HSCs. However, a number of microvilli projected into the intercellular clefts on the lateral aspects of the hepatocytes, features typical of a growth phase. Microarray profiles indicated that ∼82% of 16 605 probes were within a twofold range difference between h-heps and c-heps. Cluster and principal component analyses showed that the gene expression patterns of c-heps were extremely similar to those of h-heps. In conclusion, the chimeric mouse livers were normally reconstructed with h-heps and m-HSCs, and expressed most human genes at levels similar to those in human livers, although the chimeric livers showed morphological characteristics typical of growth.

Evaluation of the human relevance of the constitutive androstane receptor-mediated mode of action for rat hepatocellular tumor formation by the synthetic pyrethroid momfluorothrin

“…Recently, mice with human hepatocyte chimeric livers have been produced by transplanting human hepatocytes into albumin enhancer/promoter-driven urokinasetype plasminogen activator-transgenic severe combined immunodeficient (uPA/SCID) mice (Tateno et al, 2015a(Tateno et al, , 2015b(Tateno et al, , 2011(Tateno et al, , 2004(Tateno et al, , 2013. The host mouse hepatocytes are replaced with human hepatocytes in the livers of the chimeric mice.…”

“…Chimeric mice with human hepatocytes (PXB mouse®, PhoenixBio Co., Ltd.) were produced as previously described (Tateno et al, 2013(Tateno et al, , 2004. Briefly, cryopreserved human hepatocytes from donors BD85, BD87 and BD195 were purchased from BD Biosciences, Woburn, MA, USA.…”

Identification of potential genomic biomarkers for Sjögren’s syndrome using data pooling of gene expression microarrays

“…Eight datasets were downloaded from GEO, and one dataset was downloaded from ArrayExpress. We also downloaded the related published studies [11][12][13][14][15][16][17]. These datasets are summarized in Table 1.…”

Validation of uPA/SCID Mouse with Humanized Liver as a Human Liver Model: Protein Quantification of Transporters, Cytochromes P450, and UDP-Glucuronosyltransferases by LC-MS/MS

“…A similar range of difference was found in liver of PXB mice in this study, though in cultured human hepatocytes, the protein expression levels of transporters tended to be greater than those in human liver. Since humanized liver retains the structure of human liver tissue (Tateno et al, 2013), our results suggest that humanized liver of PXB mice is a useful model for analyzing human hepatic drug metabolism in vivo.…”