Isolation, Characterization, and Transplantation of Bone Marrow-Derived Hepatocyte Stem Cells

Avital, Itzhak; Inderbitzin, Daniel; Aoki, Takeshi; Tyan, Dolly B.; Cohen, Arthur H.; Ferraresso, Chiara; Rózga, Jacek; Arnaout, Walid S.; Demetriou, Achilles A.

doi:10.1006/bbrc.2001.5712

Cited by 187 publications

(151 citation statements)

References 21 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. Human umbilical cord blood has also been suggested to be a reliable source of transplantable hepatic progenitor cells (15), and some limited attempts have been made to demonstrate that these cells can differentiate without cellular fusion (28).…”

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%

“…The canals of Hering contain bipotential liver cell progenitors that can proliferate in response to severe liver injury along hepatocyte or biliary epithelial differentiation pathways, particularly if the normal proliferative potential of hepatocytes is blocked prior to a regenerative stimulus [5,6]. After bone marrow transplantation, hepatocytes and biliary cells of donor marrow origin are detectable within the liver in animals and man [7][8][9]. After liver transplantation between non-gender-identical humans, hepatocytes that must have derived from extrahepatic sources are recognized [6,10].…”

Section: Introductionmentioning

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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“…Several putative mechanisms for the emergence of improved liver function after bone marThe finding of improvement in ascites in a significant number of the patients is impressive and somewhat surrow-derived stem cell transplantation should be consid- ered. First, bone marrow could theoretically harbor a (4,16,29,30). Fourth, bone marrow-derived stem cells could be plastic and home to the injured liver and differspecialized endodermal stem cell capable of producing hepatocytes and other epithelial cells.…”

Section: Methodsmentioning

confidence: 99%

Autologous Hematopoietic Stem Cell Transplantation in 48 Patients with End-Stage Chronic Liver Diseases

Zekri²,

et al. 2010

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The only presently viable treatment for end-stage liver disease is whole organ transplantation. However, there are insufficient livers available. The aim of the present study is to provide autologous bone marrowderived stem cells as a potential therapeutic for patients with end-stage cirrhosis. This is a retrospective chart review of autologous stem cell treatment in 48 patients, 36 with chronic end-stage hepatitis C-induced liver disease and 12 with end-stage autoimmune liver disease. For all patients, granulocyte colony-stimulating factor was administered to mobilize their hematopoietic stem cells. Following leukapheresis, CD34 + stem cells were isolated, amplified, and partially differentiated in culture, then reinjected into each subject via their hepatic artery or portal vein. Treatment was generally well tolerated with the expected moderate but transient bone pain from G-CSF in less than half of the patients. Three patients had serious treatment-related complications, and only 20.8% of these end-stage liver disease patients died during 12 months of follow up. For all patients there was a statistically significant decrease in ascites. There was clinical and biochemical improvement in a large percentage of patients who received the transplantation. In the viral group, there were marked changes in albumin (p = 0.0003), bilirubin (p = 0.04), INR (p = 0.0003), and ALT levels (p = 0.02). In the autoimmune group, values also improved significantly for albumin (p = 0.001), bilirubin (p = 0.002), INR (p = .0005), and ALT levels (p = 0.003). These results suggest that autologous CD34 + stem cell transplantation may be safely administered and appears to offer some therapeutic benefit to patients with both viral and autoimmune-induced end-stage liver disease.

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Isolation, Characterization, and Transplantation of Bone Marrow-Derived Hepatocyte Stem Cells

Cited by 187 publications

References 21 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

Autologous Hematopoietic Stem Cell Transplantation in 48 Patients with End-Stage Chronic Liver Diseases

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