Liver‐Specific Gene Expression in Cultured Human Hematopoietic Stem Cells

Fiegel, Henning C.; Lioznov, Michael; Cortes-Dericks, Lourdes; Lange, Claudia; Kluth, Dietrich; Zander, Axel R.

doi:10.1634/stemcells.21-1-98

Cited by 110 publications

(64 citation statements)

References 33 publications

(44 reference statements)

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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%

“…Studies identifying potential stem cells with hepatic markers derived from bone marrow described by Fiegel et al showed albumin and CK-19 expression by RT-PCR only after culture in a collagen matrix for 35 days [18], and Jiang et al also noted the differentiated phenotype after many weeks in culture [32].…”

Section: C) Cells Expressed Cytokeratins Suggestive Of Hepatocytes (Cmentioning

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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“…16 To investigate whether MBL is synthesised by CD34 þ cells, mononuclear cells were obtained from three BM samples (donors with MBL wild type) and CD34 þ cells were separated and cultured as described earlier. 17 MBL concentrations in cultures were analysed on day 14 using the ELISA.…”

Section: Patientsmentioning

confidence: 99%

Influence of mannose-binding lectin genotypes and serostatus in allo-SCT: analysis of 131 recipients and donors

Neth

Bacher

Das

et al. 2009

Bone Marrow Transplant

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Mannan-binding lectin (MBL) deficiency is determined by MBL gene polymorphisms and is associated with an increased infection risk. To clarify the role of MBL in Allo-SCT, 131 recipients-donors were analysed. MBL genotypes were determined by PCR and heteroduplex analyses, MBL serum levels by ELISA, and MBL oligomers by western blotting. MBL levels o400 ng/ml were associated with increased susceptibility to fungal pneumonia (7/12 vs 35/111; P ¼ 0.04, adjusted P ¼ 0.002), HSV/VZV (7/12 vs 26/111; P ¼ 0.03), CMV reactivation and acute GVHD. Donor genotypes had no influence. Pre-SCT MBL levels corresponded to recipients' genotypes (Po0.001), changed significantly post-SCT, but were not influenced by donors' genotypes. MBL oligomer profiles were similar pre-/post-SCT. Cultured CD34 þ cells were found not to synthesise MBL. In conclusion, low MBL levels pre-transplant predisposed patients to sepsis, fungal and viral infection. Donors' MBL genotypes did not influence infection rates. Prospective studies should clarify the importance of MBL as a prelude for MBL replacement after SCT.

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Liver‐Specific Gene Expression in Cultured Human Hematopoietic Stem Cells

Abstract: ABSTRACT

Cited by 110 publications

References 33 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

Influence of mannose-binding lectin genotypes and serostatus in allo-SCT: analysis of 131 recipients and donors

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