Hepatocyte Growth Factor Induces Differentiation of Adult Rat Bone Marrow Cells into a Hepatocyte Lineage in Vitro

Oh, Seh Hoon; Miyazaki, Masahiro; Kouchi, Hirosuke; Inoue, Yusuke; Sakaguchi, Masakiyo; Tsuji, Toshiya; Shima, Nobuyuki; Higashio, Kanji; Namba, Masayoshi

doi:10.1006/bbrc.2000.3985

Cited by 166 publications

(128 citation statements)

References 27 publications

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“…There have been a number of studies suggesting that bone marrow-marrow progenitor cells may differentiate into hepatocyte-like cells in rodents (18)(19)(20)(21)(22). Studies utilizing human bone marrow-derived cells have produced similar results both in vitro and in animal studies (17,(23)(24)(25); however, the phenotype of the hepatocyte-like cells has not been adequately defined in most cases, and the demonstration of cellular fusion (26,27) posed a major limitation to the interpretation of animal transplantation studies.…”

Section: Discussionmentioning

confidence: 99%

Side Population Cells Derived from Adult Human Liver Generate Hepatocyte-like Cells In Vitro

et al. 2005

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We sought to determine whether hepatic side population (SP) cells derived from adult human liver possess the potential of a novel candidate hepatic stem cell. Human cadaveric donor liver was subjected to collagenase perfusion and hepatocytes were separated from nonparenchymal cells by differential centrifugation. SP cells were isolated from the nonparenchymal portion after Hoechst 33342 staining. Since CD45 is a panleukocyte antigen, CD45-negative SP cells were separated from the vast majority of CD45-positive SP cells (90%), and hepatic growth medium was used to culture both groups. Both CD45-negative and CD45-positive hepatic SP cells generated colonies in the hepatic growth medium in 2-3 weeks. The colonies yielded large cells morphologically consistent with human hepatocytes, demonstrating granule-rich cytoplasm, dense, often double nuclei, and intracellular lipofuscin pigment. The cultured cells from both sources were positive for markers of human hepatocytes: HepPar, cytokeratin 8 (CK8), and human albumin. Reverse transcriptasepolymerase chain reaction (RT-PCR) performed on both groups demonstrated positivity for additional liver markers including human albumin, CK18, α-1 anti-trypsin, and the human cytochrome P450 enzyme CYP2B6. Double immunostaining (CD45 and HepPar) and RT-PCR confirmed that the hepatocyte-like cells derived from the CD45-negative SP cells acquired HepPar positivity but had no detectable CD45 antigen expression. In contrast, the cultured cells derived from the CD45-positive SP cells also acquired HepPar positivity, but only a minimal fraction expressed the CD45 antigen. We conclude that hepatic SP cells derived from the nonparenchymal portion of human liver are a potential source of human hepatocytes irrespective of their CD45 status, and further animal studies will be required to assess their regenerative potential. Keywordshuman; side population; stem cells; hepatic; CD45 SP cells are identified by flow cytometry because of their unique ability to efflux Hoechst 33342 dye and have been shown in animal studies to be highly enriched stem cells (1,2). While the true adult liver stem cell has not been characterized, human hepatic side population (SP) cells might contain certain cells that might have hepatic progenitor-like properties. The liver in a normal healthy adult maintains a balance between cell gain and cell loss. It is composed mainly of two epithelial cell types, hepatocytes and bile ductular cells, both of which have high regenerative capacity. After hepatic injury, a regenerative process repairs the liver and brings it back to its original mass within 10 days (3). This restoration of mild to moderate cell loss and "wear and tear" renewal is largely achieved by hepatocyte self-replication. A more severe liver injury activates the endogenous stem cell population; these are often termed "oval" cells and are much smaller than mature hepatocytes (4). These oval cells display a distinct phenotype and have been shown to be a bipotential progenitor of two types of cells found ...

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Section: Discussionmentioning

confidence: 99%

Side Population Cells Derived from Adult Human Liver Generate Hepatocyte-like Cells In Vitro

et al. 2005

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“…Studies have been undertaken mostly in rodents [7,9,[14][15][16][17], but in humans, bone marrow [8,[18][19][20], amniotic tissue [21], cord blood [22], and adult and fetal liver have been studied.…”

Section: Discussionmentioning

confidence: 99%

Epithelial Colonies Cultured from Human Explanted Liver in Subacute Hepatic Failure Exhibit Hepatocyte, Biliary Epithelial, and Stem Cell Phenotypic Markers

et al. 2003

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The liver in subacute hepatic failure may become enriched for hepatic progenitor cells. Liver tissue from such a patient was collagenase digested and, from the nonparenchymal cell fraction, epithelioid colonies were developed. Albumin and alpha-1-antitrypsin (AAT) were secreted for greater than 120 days from these colonies. Reverse transcription-polymerase chain reaction showed expression of markers of both hepatocyte and biliary epithelial phenotypes (cytokeratins 7, 18, and 19, albumin and AAT, hepatocyte growth factor receptor, transforming growth factor beta receptor type II, gamma-glutamyl transpeptidase, biliary glycoprotein). The cell cycle regulator p21 was also expressed. The POU domain transcription factor octamer-binding protein 4 was present in these cells, but not in RNA or cDNA prepared from adult human liver. These markers were maintained even after 165 days culture. Proliferating epithelial-like cells with combined hepatocyte-and biliary-epithelial-specific functional markers and a stem cell marker can be isolated from the nonparenchymal fraction of liver cells in subacute hepatic failure. Stem

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“…Two days after the culture, in the differentiation medium, EBs were plated on tissue culture plates coated with 0.1% gelatin and allowed to attach and spread in further culture. In some experiments, several growth factors were added to culture media for hepatic maturation at the following concentrations: 20 ng/mL recombinant human acidic fibroblast growth factor; 10 ng/mL recombinant human basic fibroblast growth factor (bFGF) 18 (Gibco-BRL); 10 ng/mL rat recombinant hepatocyte growth factor (HGF) [19][20][21] (Toyobo Co., Ltd. Osaka, Japan); 10 ng/mL recombinant mouse oncostatin M 22 (R&D Systems, Minneapolis, MN); 10 -7 mol/L dexamethasone (Sigma); ITS (insulin 10 g/mL, transferrin 5 g/mL, selenium 5 ng/mL) (Gibco-BRL). To induce hepatic maturation of EBs in vitro, we added several growth factors to culture media.…”

Section: Methodsmentioning

confidence: 99%

Embryoid-body cells derived from a mouse embryonic stem cell line show differentiation into functional hepatocytes

Tanaka²,

et al. 2002

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Embryonic stem (ES) cells have a potential to differentiate into various progenitor cells.Here we investigated the differentiation capacity of mouse ES cells into hepatocytes both in vitro and in vivo. During the culture of embryoid bodies (EBs) derived from ES cells, albumin (ALB) messenger RNA (mRNA) was expressed within 12 days after removal of leukemia inhibitory factor, and ␣-fetoprotein (AFP) mRNA was observed within 9 days without additional exogenous growth factors. In ES cells and early EBs, by contrast, neither ALB mRNA nor AFP mRNA was observed. ALB protein was first detected at day 15 and the level increased with the culture period. The differentiation of EBs facilitated the synthesis of urea with the culture period, whereas early EBs and ES cells produced no urea. These results suggest that cultured EBs contain hepatocytes capable of producing ALB and urea. ES cells and the isolated cells from EBs were transplanted through portal vein to the liver after 30% partial hepatectomy of female mice pretreated with 2-acetylaminofluorene. Four weeks after transplantation with isolated cells from day-9 EBs, ES-derived cells containing Y-chromosome in the liver were positive for ALB (0.2% of total liver cells), whereas teratoma was found in mice transplanted with ES cells or EBs up to day 6. The incidence of teratoma was decreased with the culture duration and no teratoma was observed in the liver transplanted with isolated cells from day-9 EBs. In conclusion, our in vitro and in vivo experiments revealed that cultured EBs contain functional hepatocytes or hepatocyte-like cells. (HEPATOLOGY 2002;36:22-29.)

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Hepatocyte Growth Factor Induces Differentiation of Adult Rat Bone Marrow Cells into a Hepatocyte Lineage in Vitro

Cited by 166 publications

References 27 publications

Side Population Cells Derived from Adult Human Liver Generate Hepatocyte-like Cells In Vitro

Side Population Cells Derived from Adult Human Liver Generate Hepatocyte-like Cells In Vitro

Epithelial Colonies Cultured from Human Explanted Liver in Subacute Hepatic Failure Exhibit Hepatocyte, Biliary Epithelial, and Stem Cell Phenotypic Markers

Embryoid-body cells derived from a mouse embryonic stem cell line show differentiation into functional hepatocytes

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