title: 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.
authors.affiliation: PhoenixBio Co., Ltd., Higashihiroshima, Japan.
authors.forename: Chise
authors.initials: C
authors.lastname: Tateno
authors.forename: Fuyuki
authors.initials: F
authors.lastname: Miya
authors.forename: Kenjiro
authors.initials: K
authors.lastname: Wake
authors.forename: Miho
authors.initials: M
authors.lastname: Kataoka
authors.forename: Yuji
authors.initials: Y
authors.lastname: Ishida
authors.forename: Chihiro
authors.initials: C
authors.lastname: Yamasaki
authors.forename: Ami
authors.initials: A
authors.lastname: Yanagi
authors.forename: Masakazu
authors.initials: M
authors.lastname: Kakuni
authors.forename: Eddie
authors.initials: E
authors.lastname: Wisse
authors.forename: Fons
authors.initials: F
authors.lastname: Verheyen
authors.forename: Kouji
authors.initials: K
authors.lastname: Inoue
authors.forename: Kota
authors.initials: K
authors.lastname: Sato
authors.forename: Atsushi
authors.initials: A
authors.lastname: Kudo
authors.forename: Shigeki
authors.initials: S
authors.lastname: Arii
authors.forename: Toshiyuki
authors.initials: T
authors.lastname: Itamoto
authors.forename: Toshimasa
authors.initials: T
authors.lastname: Asahara
authors.forename: Tatsuhiko
authors.initials: T
authors.lastname: Tsunoda
authors.forename: Katsutoshi
authors.initials: K
authors.lastname: Yoshizato
mesh_terms: D000704:Analysis of Variance; D000818:Animals; D002448:Cell Adhesion / Q000502:physiology; D015687:Cell Hypoxia / Q000502:physiology; D005260:Female; D020869:Gene Expression Profiling; D055166:Hepatic Stellate Cells / Q000166:cytology; D022781:Hepatocytes / Q000166:cytology*; D006801:Humans; D007150:Immunohistochemistry; D007728:Kupffer Cells / Q000166:cytology; D008099:Liver / Q000737:chemistry / Q000166:cytology*; D008297:Male; D051379:Mice; D016513:Mice, SCID; D046888:Tissue Array Analysis / Q000379:methods; D014183:Transplantation, Heterologous