A Lineage of Myeloid Cells Independent of Myb and Hematopoietic Stem Cells

Science ◽  
2012 ◽  
Vol 336 (6077) ◽  
pp. 86-90 ◽  
Author(s):  
Christian Schulz ◽  
Elisa Gomez Perdiguero ◽  
Laurent Chorro ◽  
Heather Szabo-Rogers ◽  
Nicolas Cagnard ◽  
...  
Keyword(s):  
Stem Cells ◽  
Transcription Factor ◽  
Hematopoietic Stem Cells ◽  
Langerhans Cells ◽  
Cell Populations ◽  
Hematopoietic Stem ◽  
Microglia Cell ◽  
Adult Mice ◽  
Monocytes And Macrophages ◽  
Epidermal Langerhans Cells

Macrophages and dendritic cells (DCs) are key components of cellular immunity and are thought to originate and renew from hematopoietic stem cells (HSCs). However, some macrophages develop in the embryo before the appearance of definitive HSCs. We thus reinvestigated macrophage development. We found that the transcription factor Myb was required for development of HSCs and all CD11bhighmonocytes and macrophages, but was dispensable for yolk sac (YS) macrophages and for the development of YS-derived F4/80brightmacrophages in several tissues, such as liver Kupffer cells, epidermal Langerhans cells, and microglia—cell populations that all can persist in adult mice independently of HSCs. These results define a lineage of tissue macrophages that derive from the YS and are genetically distinct from HSC progeny.

Reversible Expression of CD34 by Murine Hematopoietic Stem Cells

Blood ◽  
1999 ◽  
Vol 94 (8) ◽  
pp. 2548-2554 ◽  
Author(s):  
Takashi Sato ◽  
Joseph H. Laver ◽  
Makio Ogawa
Keyword(s):  
Stem Cells ◽  
Hematopoietic Stem Cells ◽  
Bone Marrow Cells ◽  
Cell Populations ◽  
Hematopoietic Stem ◽  
Adult Mice ◽  
Cd34 Expression ◽  
Marrow Cells

We used a mouse transplantation model to address the recent controversy about CD34 expression by hematopoietic stem cells. Cells from Ly-5.1 C57BL/6 mice were used as donor cells and Ly-5.2 mice were the recipients. The test cells were transplanted together with compromised marrow cells of Ly-5.2 mice. First, we confirmed that the majority of the stem cells with long-term engraftment capabilities of normal adult mice are CD34−. We then observed that, after the injection of 150 mg/kg 5-fluorouracil (5-FU), stem cells may be found in both CD34− and CD34+ cell populations. These results indicated that activated stem cells express CD34. We tested this hypothesis also by using in vitro expansion with interleukin-11 and steel factor of lineage−c-kit+ Sca-1+ CD34− bone marrow cells of normal mice. When the cells expanded for 1 week were separated into CD34− and CD34+ cell populations and tested for their engraftment capabilities, only CD34+ cells were capable of 2 to 5 months of engraftment. Finally, we tested reversion of CD34+ stem cells to CD34− state. We transplanted Ly-5.1 CD34+post–5-FU marrow cells into Ly-5.2 primary recipients and, after the marrow achieved steady state, tested the Ly-5.1 cells of the primary recipients for their engraftment capabilities in Ly-5.2 secondary recipients. The majority of the Ly-5.1 stem cells with long-term engraftment capability were in the CD34− cell fraction, indicating the reversion of CD34+ to CD34−stem cells. These observations clearly demonstrated that CD34 expression reflects the activation state of hematopoietic stem cells and that this is reversible.

Dissociation of bone resorption and bone formation in adult mice transplanted with OC/OC hematopoietic stem cells

Bone ◽  
2010 ◽  
Vol 47 ◽  
pp. S139
Author(s):  
C. Flores ◽  
C.S. Thudium ◽  
G. Langenbach ◽  
M. Brüel ◽  
N.A. Sims ◽  
...  
Keyword(s):  
Stem Cells ◽  
Bone Resorption ◽  
Bone Formation ◽  
Hematopoietic Stem Cells ◽  
Hematopoietic Stem ◽  
Adult Mice

scholarly journals A Novel, Myeloid Transcription Factor, C/EBPε, Is Upregulated During Granulocytic, But Not Monocytic, Differentiation

Blood ◽  
1997 ◽  
Vol 90 (7) ◽  
pp. 2591-2600 ◽  
Author(s):  
Roberta Morosetti ◽  
Dorothy J. Park ◽  
Alexey M. Chumakov ◽  
Isabelle Grillier ◽  
Masaaki Shiohara ◽  
...  
Keyword(s):  
Stem Cells ◽  
Transcription Factor ◽  
Hematopoietic Stem Cells ◽  
Cell Line ◽  
Cell Lines ◽  
Myeloid Leukemia ◽  
Leukemia Cell ◽  
Rt Pcr ◽  
Hematopoietic Stem ◽  
Nb4 Cells

Human C/EBPε is a newly cloned CCAAT/enhancer-binding transcription factor. Initial studies indicated it may be an important regulator of human myelopoiesis. To elucidate the range of expression of C/EBPε, we used reverse transcription-polymerase chain reaction (RT-PCR) analysis and examined its expression in 28 hematopoietic and 14 nonhematopoietic cell lines, 16 fresh myeloid leukemia samples, and normal human hematopoietic stem cells and their mature progeny. Prominent expression of C/EBPε mRNA occurred in the late myeloblastic and promyelocytic cell lines (NB4, HL60, GFD8), the myelomonoblastic cell lines (U937 and THP-1), the early myeloblast cell lines (ML1, KCL22, MDS92), and the T-cell lymphoblastic leukemia cell lines CEM and HSB-2. For the acute promyelocytic leukemia cell line NB4, C/EBPε was the only C/EBP family member that was easily detected by RT-PCR. No C/EBPε mRNA was found in erythroid, megakaryocyte, basophil, B lymphoid, or nonhematopoietic cell lines. Most acute myeloid leukemia samples (11 of 12) from patients expressed C/EBPε. Northern blot and RT-PCR analyses showed that C/EBPε mRNA decreased when the HL60 and KG-1 myeloblast cell lines were induced to differentiate toward macrophages. Similarly, Western blot analysis showed that expression of C/EBPε protein was either unchanged or decreased slightly as the promyelocytic cell line NB4 differentiated down the macrophage-like pathway after treatment with a potent vitamin D3 analog (KH1060). In contrast, C/EBPε protein levels increased dramatically as NB4 cells were induced to differentiate down the granulocytic pathway after exposure to 9-cis retinoic acid. Furthermore, very early, normal hematopoietic stem cells (CD34+/CD38−), purified from humans had very weak expression of C/EBPε mRNA, but levels increased as these cells differentiated towards granulocytes. Likewise, purified granulocytes appeared to express higher levels of C/EBPε mRNA than purified macrophages. Addition of phosphothiolated antisense, but not sense oligonucleotides to C/EBPε, decreased clonal growth of HL-60 and NB4 cells by about 50% compared with control cultures. Taken together, our results indicate that expression of C/EBPε is restricted to hematopoietic tissues, especially myeloid cells as they differentiate towards granulocytes and inhibition of its expression in HL-60 and NB4 myeloblasts and promyelocytes decreased their proliferative capacity. Therefore, this transcriptional factor may play an important role in the process of normal myeloid development.

The transcription factor Erg is essential for definitive hematopoiesis and the function of adult hematopoietic stem cells

2008 ◽  
Vol 9 (7) ◽  
pp. 810-819 ◽  
Author(s):  
Stephen J Loughran ◽  
Elizabeth A Kruse ◽  
Douglas F Hacking ◽  
Carolyn A de Graaf ◽  
Craig D Hyland ◽  
...  
Keyword(s):  
Stem Cells ◽  
Transcription Factor ◽  
Hematopoietic Stem Cells ◽  
Hematopoietic Stem ◽  
Definitive Hematopoiesis

scholarly journals The transcription factor Foxm1 is essential for the quiescence and maintenance of hematopoietic stem cells

2015 ◽  
Vol 16 (8) ◽  
pp. 810-818 ◽  
Author(s):  
Yu Hou ◽  
Wen Li ◽  
Yue Sheng ◽  
Liping Li ◽  
Yong Huang ◽  
...  
Keyword(s):  
Stem Cells ◽  
Transcription Factor ◽  
Hematopoietic Stem Cells ◽  
Hematopoietic Stem

Role of the Transcription Factor LYL-1 in the Adult and Fetal Hematopoietic Stem Cells.

Blood ◽  
2004 ◽  
Vol 104 (11) ◽  
pp. 2775-2775
Author(s):  
Claude Capron ◽  
Catherine Lacout ◽  
Yann Lecluse ◽  
Isabelle Poullion ◽  
Fedor Svinarchouk ◽  
...  
Keyword(s):  
Stem Cells ◽  
Transcription Factor ◽  
Hematopoietic Stem Cells ◽  
Absolute Number ◽  
Binding Motif ◽  
Hematopoietic Stem ◽  
Cell Counts ◽  
Significant Difference ◽  
The Absolute

Abstract The hematopoietic stem cells (HSC) have the ability to self-renew and to give rise to all blood lineages. These processes occur via a hierarchy of progenitors with progressively more limited differentiation and self-renewal potential and are orchestrated by specialized protein such as transcription factors. LYL-1 protein contains a basic helix-loop-helix DNA binding motif also found in several proteins involved in the control of cellular proliferation and differentiation such as SCL/TAL-1. As LYL-1 shares an 80% homology at the protein level with SCL/TAL-1, we wanted to determine the function of LYL-1 in hematopoiesis and particularly on HSC. For this study, we used knock in lyl-1−/− mice in which exon 4 was replaced by LacZ/Neo cassette. Lyl−/− mice are viable and have normal blood cell counts as well as a normal marrow cellularity. In addition, using a hematopoietic colony forming cells (CFCs) assay, no significant difference was seen in the myeloid CFCs of either lyl-1−/− or lyl-1+/+ BM and FL cells except a 2-fold increase in the absolute number of BFU-E in lyl-1−/− FL as compared to lyl-1+/+ FL. We analyzed more primitive progenitors in details because using Fluorecein Di-beta Galactopyranoside (FDG)-staining assay, we showed that lyl-1 is mainly expressed in primitive Lin− Sca-1+ c-Kit+ cells (LSK) cells from either BM or FL (91 ± 7% and 78 ± 5% of FDG positive cells in lyl-1−/− BM and FL LSK cells, respectively). In addition, analysis of lyl-1−/− and lyl-1+/+ cells revealed a 1.8-fold and 2-fold decrease in the percentage of primitive LSK in BM and FL, respectively, as compared to wild type cells. Furthermore, using the Hoechst 33342 efflux assay, we noticed a significant decrease in the absolute number of more primitive LSK-SP (side population) cells in lyl-1−/− BM as compared to lyl-1+/+ BM cells (52800 ± 5412 cells/femur versus 91080 ± 8475 cells/femur, respectively) suggesting an important role of LYL-1 in the HSC function. In order to confirm this hypothesis, in vivo assays were performed. We observed a 1.5-fold decrease in the lyl-1−/− BM and FL day 12 CFU-S content as compared to lyl-1+/+ cells. Adoptive transfer experiments were subsequently performed using lethally irradiated Ly5.1 mice. Data showed that lyl-1−/− cells from either BM or FL displayed a hematopoietic reconstitution defect in competitive repopulation assays. Indeed, Ly5.1 recipients were injected with a mixture of 5x106 (5:1), 106 (1:1) or 0.5x106 (0.5:1) lyl-1−/− or lyl-1+/+ Ly5.2 expressing cells and 106 competitive BM Ly5.1 expressing cells. All hosts engrafted with lyl-1−/− BM cells shown a significant reduced levels of chimerism (% of circulating Ly5.2+ cells) as compared to hosts engrafted with lyl-1+/+ BM donors (4.3 ± 2.8% (5:1); 7.5 ± 5.5% (1:1); 0.6 ± 0.3% (0.5:1) in lyl-1−/− BM cells versus 66 ± 8% (5:1); 52 ± 9% (1:1); 53 ± 10% (0.5:1) in lyl-1+/+ BM cells) and similar difference was observed with FL donors (45 ± 2% (5:1); 25 ± 5% (1:1); 11 ± 5% (0.5:1) in lyl-1−/− FL cells versus 83 ± 1% (5:1); 70 ± 3% (1:1); 53 ± 6% (0.5:1) in lyl-1+/+ FL cells). This altered defect in HSC was also confirmed using LTC-IC in vitro experiments. Altogether, our results demonstrate an important role of the transcription factor LYL-1 on the maintenance of HSC properties.

Induction of Adipocyte Differentiation and Reduction of Hematopoesis-Supporting Activity of Mouse Stromal Cells by Suppression of Transcription Factor GATA-2.

Blood ◽  
2005 ◽  
Vol 106 (11) ◽  
pp. 1392-1392
Author(s):  
Yoko Okitsu ◽  
Hideo Harigae ◽  
Masanori Seki ◽  
Toru Fujiwara ◽  
Shinichiro Takahashi ◽  
...  
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Transcription Factor ◽  
Hematopoietic Stem Cells ◽  
Stromal Cells ◽  
Colony Formation ◽  
Adipocyte Differentiation ◽  
Fetal Liver ◽  
Expression Level ◽  
Hematopoietic Stem

Abstract (Introduction) Aplastic anemia (AA) is characterized by peripheral pancytopenia and fatty bone marrow. An immunological attack to hematopoietic stem cells has been thought to be responsible for the development of the disease. Previously, we reported the expression of transcription factor GATA-2 is significantly decreased in CD34 positive cells in AA. Together with the phenotypes of hematopoietic stem cells in GATA-2 hetero-knockout mice, GATA-2 down-regulation may play a role in the reduction of a stem cell pool observed in AA. On the other hand, GATA-2 has been shown to be essential for the maintenance of immaturity of preadipocytes. If a pathological immune response in AA decreases the level of GATA-2 expression in not only hematopoietic stem cells but also stromal preadipocytes, it may accelerate the maturation of preadipocytes, leading to the formation of fatty bone marrow. To explore this possibility, the phenotypic change of stromal preadipocytes by suppression of GATA-2 was examined in this study. (Method) The GATA-2 expression level was suppressed by using siRNA for GATA-2 in mouse stromal preadipocyte cell lines, TBR9 and TBR343. After the treatment with siRNA, the adipocyte differentiation was induced by the incubation with insulin and dexamethasone for 7days. Then, the maturation level was examined by oil drops formation judged by oil red staining, and by the expression level of adipcin and PPAR-γ mRNA. Supporting activity of hematopoietic colony formation was also evaluated by using mouse fetal liver cells after siRNA treatment. (Results) By using designed siRNA, the GATA-2 expression was suppressed to 30% of control, whereas the expression level of GATA-3, which is co-expressed in preadipocytes, was unchanged. When GATA-2 was suppressed by siRNA, the oil drop formation and adipocyte-specific gene expression was significantly accelerated in both of stromal cells. Furthermore, the number of fetal liver hematopoetic colonies was significantly decreased by suppression of GATA-2, suggesting that GATA-2 down-regulation in stromal preadipocytes results in not only the acceleration of the maturation but also the reduced supporting activity of hematopoietic colony formation (Conclusion) These results suggest that suppression of GATA-2 in hematopoietic tissues induces the characteristic features of AA, i.e., decreased the number of hematopoietic stem cells and increased number of mature adipocytes.

Ubiquitin Ligase Component Fbw7 Regulates the Quiescence of Hematopoietic Stem Cells.

Blood ◽  
2006 ◽  
Vol 108 (11) ◽  
pp. 79-79 ◽  
Author(s):  
Sahoko Matsuoka ◽  
Yuichi Oike ◽  
Ichiro Onoyama ◽  
Keiyo Takubo ◽  
Keisuke Ito ◽  
...  
Keyword(s):  
Stem Cells ◽  
Hematopoietic Stem Cells ◽  
Ubiquitin Ligase ◽  
Mononuclear Cells ◽  
Human Cancer ◽  
Self Renewal ◽  
Hematopoietic Stem ◽  
Human Cancer Cell Lines ◽  
Adult Mice ◽  
Deficient Mice

Abstract Fbw7 is a SCF ubiquitin ligase component that catalyzes the ubiquitination of c-Myc, Cyclin E, and Notch. In several human cancer cell lines and primary cancer cells, Fbw7 is mutated and functions as a tumor suppressor gene. Previously we have reported that Fbw7-deficient mice died at embryonic day 10.5–11.5 with deficiencies in hematopoietic and vascular development, indicating that Fbw7 has a pivotal role in hematopoiesis (Tsunematsu R et al. J Biol Chem. 2004). Fbw7 is widely expressed in various hematopoietic lineages in BM of adult mice, but little has been known about the function of Fbw7 in hematopoiesis. To assess the requirement of Fbw7 in adult hematopoietic cells, we generated Fbw7-deficient mice by conditional gene targeting. Fbw7 was conditionally deleted from Mx-1-Cre;Fbw7fl/− adult mice by injection of pIpC over 1 week to induce Cre expression. We examined Fbw7fl/+ littermates as a control. We found progressive pancytopenia in Fbw7-deficient mice. Furthermore, most Fbw7-deficient mice developed leukemia (mainly ALL) within 3 months after pIpC treatment, suggesting that Fbw7 is essential to maintain normal hematopoiesis and loss of Fbw7 accelerates leukemogenesis. The portion of Fbw7-deficient Lin−Sca-1+c-Kit+CD34− hematopoietic stem cells (HSCs) in the G0 phase was 2.5-fold decreased and the frequency of cell division of Fbw7-deficient HSCs markedly increased in culture. These data suggest that Fbw7 promotes quiescence of HSCs. To examine the function of Fbw7-deficient HSCs, we transplanted 1500 Lin−Sca-1+cKit+ BM cells from Fbw7-dificient mice or littermate controls into lethally irradiated recipient mice with 4×105 normal BM mononuclear cells. In the result, Fbw7-deficient HSCs are impaired in long-term repopulating activity and multipotency. It has been reported that c-Myc controls the self-renewal activity of HSCs through the cell adhesion to the osteoblastic niche (Wilson A et al. Genes Dev. 2004). We found that c-Myc is significantly accumulated in Fbw7-deficient Lin−Sca-1+cKit+ BM cells, suggesting that HSCs leave the niche and show the active cell cycling. We propose that a ubiquitin ligase, Fbw7 is a key mediator of HSC quiescence and self renewal capacity.

Generation of definitive hematopoietic stem cells from murine early yolk sac and paraaortic splanchnopleures by aorta-gonad-mesonephros region–derived stromal cells

Blood ◽  
2001 ◽  
Vol 98 (1) ◽  
pp. 6-12 ◽  
Author(s):  
Sahoko Matsuoka ◽  
Kohichiro Tsuji ◽  
Hiroaki Hisakawa ◽  
Ming-jiang Xu ◽  
Yasuhiro Ebihara ◽  
...  
Keyword(s):  
Stem Cells ◽  
Hematopoietic Stem Cells ◽  
Blood Vessels ◽  
Stromal Cells ◽  
Yolk Sac ◽  
Hematopoietic Stem ◽  
Adult Mice ◽  
Definitive Hematopoiesis

Abstract There is controversy as to whether murine definitive hematopoiesis originates from yolk sac (YS) or the intraembryonic region. This study reports the generation of definitive hematopoietic stem cells (HSCs) from both early YS and intraembryonic paraaortic splanchnopleures (P-Sp) on AGM-S3 stromal cells derived from the aorta-gonad-mesonephros (AGM) region at 10.5 days post coitum (dpc). YS and P-Sp cells at 8.5 dpc generated no definitive hematopoiesis-derived colony-forming cells in cocultures with AGM-S3 cells, but spleen colony-forming cells and HSCs capable of reconstituting definitive hematopoiesis in adult mice simultaneously appeared on day 4 of coculture. Precursors for definitive HSCs were present in YS and P-Sp at 8.0 dpc, a time when YS and embryo were not connected by blood vessels. It is proposed that precursors with the potential to generate definitive HSCs appear independently in YS and intraembryonic P-Sp and that the P-Sp or AGM region affords the microenvironment that facilitates generation of definitive hematopoiesis from precursors.

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