Bone-marrow derived hematopoietic stem/progenitor cells express multiple isoforms of NADPH oxidase and produce constitutively reactive oxygen species

Abstract Exposure to electromagnetic fields (EMF) has been associated with the increased risk of childhood leukemia, which arises from mutations induced within hematopoietic stem cells often through preleukemic fusion genes (PFG). In this study we investigated whether exposure to microwaves (MW) emitted by mobile phones could induce various biochemical markers of cellular damage including reactive oxygen species (ROS), DNA single and double strand breaks, PFG, and apoptosis in umbilical cord blood (UCB) cells including CD34+ hematopoietic stem/progenitor cells. UCB cells were exposed to MW pulsed signals from GSM900/UMTS test-mobile phone and ROS, apoptosis, DNA damage, and PFG were analyzed using flow cytometry, automated fluorescent microscopy, imaging flow cytometry, comet assay, and RT-qPCR. In general, no persisting difference in DNA damage, PFG and apoptosis between exposed and sham-exposed samples was detected. However, we found increased ROS level after 1 h of UMTS exposure that was not evident 3 h post-exposure. We also found that the level of ROS rise with the higher degree of cellular differentiation. Our data show that UCB cells exposed to pulsed MW developed transient increase in ROS that did not result in sustained DNA damage and apoptosis.

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FGF-2 expands murine hematopoietic stem and progenitor cells via proliferation of stromal cells, c-Kit activation, and CXCL12 down-regulation

Blood ◽

10.1182/blood-2011-11-394692 ◽

2012 ◽

Vol 120 (9) ◽

pp. 1843-1855 ◽

Cited By ~ 66

Author(s):

Tomer Itkin ◽

Aya Ludin ◽

Ben Gradus ◽

Shiri Gur-Cohen ◽

Alexander Kalinkovich ◽

...

Keyword(s):

Reactive Oxygen Species ◽

Progenitor Cells ◽

Stromal Cells ◽

Reactive Oxygen ◽

Receptor Activation ◽

Oxygen Species ◽

Hematopoietic Stem ◽

Cell Engraftment ◽

Stem And Progenitor Cells

Abstract Cytokine-induced expansion of hematopoietic stem and progenitor cells (HSPCs) is not fully understood. In the present study, we show that whereas steady-state hematopoiesis is normal in basic fibroblast growth factor (FGF-2)–knockout mice, parathyroid hormone stimulation and myeloablative treatments failed to induce normal HSPC proliferation and recovery. In vivo FGF-2 treatment expanded stromal cells, including perivascular Nestin+ supportive stromal cells, which may facilitate HSPC expansion by increasing SCF and reducing CXCL12 via mir-31 up-regulation. FGF-2 predominantly expanded a heterogeneous population of undifferentiated HSPCs, preserving and increasing durable short- and long-term repopulation potential. Mechanistically, these effects were mediated by c-Kit receptor activation, STAT5 phosphorylation, and reduction of reactive oxygen species levels. Mice harboring defective c-Kit signaling exhibited abrogated HSPC expansion in response to FGF-2 treatment, which was accompanied by elevated reactive oxygen species levels. The results of the present study reveal a novel mechanism underlying FGF-2–mediated in vivo expansion of both HSPCs and their supportive stromal cells, which may be used to improve stem cell engraftment after clinical transplantation.

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Loss of Foxo3a In FA Mice Leads to Hematopoietic Stem Cell Exhaustion That Can Be Attenuated by Natural Anti-Oxidant Quercetin.

Blood ◽

10.1182/blood.v116.21.1166.1166 ◽

2010 ◽

Vol 116 (21) ◽

pp. 1166-1166

Author(s):

Jie Li ◽

Jared Sipple ◽

Qishen Pang

Keyword(s):

Reactive Oxygen Species ◽

Progenitor Cells ◽

Peripheral Blood ◽

Conflicts Of Interest ◽

Genetic Disorder ◽

Specific Interaction ◽

Reactive Oxygen ◽

Oxygen Species ◽

Hematopoietic Stem ◽

Anti Oxidant

Abstract Abstract 1166 Fanconi anemia (FA) is a genetic disorder characterized by genomic instability, bone marrow (BM) failure and predisposition to cancer. However, FA mouse models do not show spontaneous genetic instability. Previous study shows that FOXO3a is associated with the FA pathway through oxidative stress-specific interaction with FANCD2. To address the consequence of loss of FOXO3a function in FA hematopoiesis, we generated Foxo3a-/-Fancd2-/- and Foxo3a-/-Fancc-/- double-knockout (DKO) mice by crossing Foxo3a+/− with Fancd2+/− or Fancc+/−; mice. Reactive oxygen species are increased in low-density BM (LDBM) cells isolated from DKO mice compared to those from single KO (SKO) or wt mice. Analysis of hematologic parameters shows significantly increased number of nucleate cells and high ratio of eosinophils in peripheral blood of DKO mice. CFU assay shows more progenitor cells in peripheral blood isolated from DKO mice. Moreover, BM progenitor cells from DKO mice exhibit lower adhesion but higher migration activity, compared to those from wt or SKO mice. Consistent with this, Cdc42 pull-down assay shows lower Cdc42 activity in DKO LDBM cells than in wt or SKO cells, indicating that decreased Cdc42 may contribute to the observed aberrant adhesion and migration activities. DKO mice show significant decrease in primitive progenitor (Lin-Sca-1+c-kit+; LSK) cells, increase in BrdU+ and G1-phase LSK cells, and impaired repopulating capacity after competitive BM transplantation, which can be attenuated by the anti-oxidant Quercetin. Taken together, loss of Foxo3a in FA mice results in FA-like syndrome, which may be resulted from increased reactive oxygen species accumulation. Disclosures: No relevant conflicts of interest to declare.

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The Role ofHibiscus sabdariffaL. (Roselle) in Maintenance ofEx VivoMurine Bone Marrow-Derived Hematopoietic Stem Cells

The Scientific World JOURNAL ◽

10.1155/2014/258192 ◽

2014 ◽

Vol 2014 ◽

pp. 1-10 ◽

Cited By ~ 7

Author(s):

Zariyantey Abdul Hamid ◽

Winnie Hii Lin Lin ◽

Basma Jibril Abdalla ◽

Ong Bee Yuen ◽

Elda Surhaida Latif ◽

...

Keyword(s):

Reactive Oxygen Species ◽

Stem Cells ◽

Bone Marrow ◽

Dna Damage ◽

Ex Vivo ◽

Reactive Oxygen ◽

Oxygen Species ◽

Sod Activity ◽

Hematopoietic Stem ◽

Murine Bone Marrow

Hematopoietic stem cells- (HSCs-) based therapy requiresex vivoexpansion of HSCs prior to therapeutic use. However,ex vivoculture was reported to promote excessive production of reactive oxygen species (ROS), exposing HSCs to oxidative damage. Efforts to overcome this limitation include the use of antioxidants. In this study, the role ofHibiscus sabdariffaL. (Roselle) in maintenance of cultured murine bone marrow-derived HSCs was investigated. Aqueous extract of Roselle was added at varying concentrations (0–1000 ng/mL) for 24 hours to the freshly isolated murine bone marrow cells (BMCs) cultures. Effects of Roselle on cell viability, reactive oxygen species (ROS) production, glutathione (GSH) level, superoxide dismutase (SOD) activity, and DNA damage were investigated. Roselle enhanced the survival(P<0.05)of BMCs at 500 and 1000 ng/mL, increased survival of Sca-1+cells (HSCs) at 500 ng/mL, and maintained HSCs phenotype as shown from nonremarkable changes of surface marker antigen (Sca-1) expression in all experimental groups. Roselle increased(P<0.05)the GSH level and SOD activity but the level of reactive oxygen species (ROS) was unaffected. Moreover, Roselle showed significant cellular genoprotective potency against H2O2-induced DNA damage. Conclusively, Roselle shows novel property as potential supplement and genoprotectant against oxidative damage to cultured HSCs.

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Reactive Oxygen Species Regulate Hematopoietic Stem Cell Self-Renewal, Migration and Development, As Well As Their Bone Marrow Microenvironment

Antioxidants and Redox Signaling ◽

10.1089/ars.2014.5941 ◽

2014 ◽

Vol 21 (11) ◽

pp. 1605-1619 ◽

Cited By ~ 123

Author(s):

Aya Ludin ◽

Shiri Gur-Cohen ◽

Karin Golan ◽

Kerstin B. Kaufmann ◽

Tomer Itkin ◽

...

Keyword(s):

Reactive Oxygen Species ◽

Bone Marrow ◽

Stem Cell ◽

Hematopoietic Stem Cell ◽

Reactive Oxygen ◽

Bone Marrow Microenvironment ◽

Oxygen Species ◽

Self Renewal ◽

Hematopoietic Stem ◽

Migration And Development

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Do vasculature reactive oxygen species play a role in the mobilization of bone marrow endothelial progenitor cells?

Journal of Hypertension ◽

10.1097/hjh.0b013e3282f2851a ◽

2008 ◽

Vol 26 (2) ◽

pp. 188-190 ◽

Cited By ~ 2

Author(s):

Hideyasu Kiyomoto ◽

Akira Nishiyama

Keyword(s):

Reactive Oxygen Species ◽

Bone Marrow ◽

Progenitor Cells ◽

Endothelial Progenitor Cells ◽

Reactive Oxygen ◽

Oxygen Species ◽

Endothelial Progenitor

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Dysfunction of endothelial progenitor cells in hyperlipidemic rats involves the increase of NADPH oxidase derived reactive oxygen species production

Canadian Journal of Physiology and Pharmacology ◽

10.1139/cjpp-2016-0142 ◽

2017 ◽

Vol 95 (5) ◽

pp. 474-480 ◽

Cited By ~ 5

Author(s):

Ting-Bo Li ◽

Jie-Jie Zhang ◽

Bin Liu ◽

Xiu-Ju Luo ◽

Qi-Lin Ma ◽

...

Keyword(s):

Reactive Oxygen Species ◽

Nadph Oxidase ◽

Progenitor Cells ◽

Endothelial Progenitor Cells ◽

Plasma Lipids ◽

Reactive Oxygen ◽

The Body ◽

Ros Production ◽

Oxygen Species ◽

Endothelial Progenitor

NADPH oxidase (NOX) is a major source of reactive oxygen species (ROS) in the body and it plays a key role in mediation of oxidative injury in the cardiovascular system. The purposes of this study are to evaluate the status of NOX in endothelial progenitor cells (EPCs) of hyperlipidemic rats and to determine whether NOX-derived ROS promotes the dysfunction of EPCs. The rats were fed on a high-fat diet for 8 weeks to establish a hyperlipidemic rat model, which showed the increased plasma lipids and the impaired functions of circulating EPCs (including the reduced abilities in migration and adhesion) accompanied by an increase in NOX activity and ROS production. Next, EPCs were isolated from normal rats and they were treated with oxidized low-density lipoprotein (ox-LDL) (100 μg/mL) for 24 h to induce a dysfunctional model in vitro. In agreement with our findings in vivo, ox-LDL treatment increased the dysfunctions of EPCs concomitant with an increase in NOX activity and ROS production; these phenomena were reversed by the NOX inhibitor. Based on these observations, we conclude that NOX-derived ROS involved in the dysfunctions of circulating EPCs in hyperlipidemic rats and inhibition of NOX might provide a novel strategy to improve EPC functions in hyperlipidemia.

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Reactive Oxygen Species Might Have Important Roles On Angiogenesis of Endothelial Progenitor Cells Derived From HCB(Human Cord Blood) Which Are Stimulated by VEGF.

Blood ◽

10.1182/blood.v114.22.1445.1445 ◽

2009 ◽

Vol 114 (22) ◽

pp. 1445-1445

Author(s):

Eun-Sun Yoo ◽

Yeung-Chul Mun ◽

Kyoung-Eun Lee ◽

Eunmi Nam ◽

Jee-Young Ahn ◽

...

Keyword(s):

Reactive Oxygen Species ◽

Nadph Oxidase ◽

Progenitor Cells ◽

Endothelial Progenitor Cells ◽

Reactive Oxygen ◽

Tube Formation ◽

Control Group ◽

Oxygen Species ◽

Endothelial Progenitor

Abstract Abstract 1445 Poster Board I-468 Purpose VEGF is a key angiogenic growth factor stimulating proliferation, migration, and tube formation on endothelial cells (ECs), that works through the VEGF receptor type 2 (VEGR2, KDR/Flk1). Reactive oxygen species (ROS) such as superoxide and H2O2 have roles signaling for molecules on angiogenesis. In present study, the aim is to investigate the roles of reactive oxygen species on neovascularization in endothelial progenitor cells. Methods Mononuclear cells isolated from UCB were cultured using EGM-2 medium with VEGF, IGF-1 and FGF or basal medium in presence or absence of VEGF for 7 days. Outgrowing endothelail progenitor cells (Yoo et al, STEM CELLS 21:228-235, 2003) at first week of culture were analyzed ROS production by dichlorofluorescein (DCF) fluorescence by use of 2,7-dichlorodihydro-fluoresceine-diacetate (H2DCF-DA). In order to determine that ROS production might involve in EPC proliferation and migration, we had analyzed the impact of N-acetyl-L-cysteine (NAC), broad spectrum ROS scavenger, and NAD(P)H oxidase inhibitor, diphenylene iodonium (DPI) using the proliferation, in vitro tube formation matrigel assay, migration assay with SDF-1/VEGF. We also analyzed the expression of NOX2-based NADPH oxidase (gp91phox) and activation of ERK2 and Akt (Thr308 and Ser473) using VEGF with or without DPI. Results Intracellular ROS level were increased during endothelial progenitor cell culture and were higher in UCB compared to that of BM and increased by VEGF treatment. Proliferation, in vitro tube formation matrigel assay and migration assay on endothelial progenitor cells using SDF-1/VEGF were decreased with additions of ROS scavenger DPI when compared with that of control group. In western blot data, NOX2-based NADPH oxidase(gp91phox) was increased by VEGF and decreased by addition of DPI. VEGF induced pERK2 expression was also decreased by DPI and that finding was correlated on down-regulation of endothelial cell proliferation by DPI. Activation of Ser473 Akt was found in control group and decreased by VEGF and rebounded by VEGF and DPI. But Thr308 Akt was not activated in our experiments. Conclusions These results suggested that NOX2-based NADPH oxidase(gp91phox)-induced ROS might play important roles on EPCs migration and proliferation by VEGF. Namely, manipulating the level of ROS biochemically may alter the pathogenesis in cardiovascular and in ischemic limb diseases. In results, these data may be useful to develop new therapeutic strategies. Disclosures No relevant conflicts of interest to declare.

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