The Cdx-Hox Pathway in Hematopoietic Stem Cell Formation from Embryonic Stem Cells

2007 ◽  
Vol 1106 (1) ◽  
pp. 197-208 ◽  
Author(s):  
C. LENGERKE ◽  
S. MCKINNEY-FREEMAN ◽  
O. NAVEIRAS ◽  
F. YATES ◽  
Y. WANG ◽  
...  
Keyword(s):  
Stem Cells ◽  
Stem Cell ◽  
Embryonic Stem Cells ◽  
Hematopoietic Stem Cell ◽  
Cell Formation ◽  
Embryonic Stem ◽  
Hematopoietic Stem

Hematopoietic progenitor/stem cells immortalized byLhx2 generate functional hematopoietic cells in vivo

Blood ◽  
2002 ◽  
Vol 99 (11) ◽  
pp. 3939-3946 ◽  
Author(s):  
Perpétua Pinto do Ó ◽  
Karin Richter ◽  
Leif Carlsson
Keyword(s):  
Stem Cells ◽  
Stem Cell ◽  
Embryonic Stem Cells ◽  
Cell Lines ◽  
Embryonic Stem ◽  
Hematopoietic Progenitor Cell ◽  
Hematopoietic Progenitor ◽  
Generate Functional ◽  
Hematopoietic Stem

Hematopoietic stem cells (HSCs) are unique in their capacity to maintain blood formation following transplantation into immunocompromised hosts. Expansion of HSCs in vitro is therefore important for many clinical applications but has met with limited success because the mechanisms regulating the self-renewal process are poorly defined. We have previously shown that expression of the LIM-homeobox gene Lhx2 in hematopoietic progenitor cells derived from embryonic stem cells differentiated in vitro generates immortalized multipotent hematopoietic progenitor cell lines. However, HSCs of early embryonic origin, including those derived from differentiated embryonic stem cells, are inefficient in engrafting adult recipients upon transplantation. To address whetherLhx2 can immortalize hematopoietic progenitor/stem cells that can engraft adult recipients, we expressed Lhx2 in hematopoietic progenitor/stem cells derived from adult bone marrow. This approach allowed for the generation of immortalized growth factor–dependent hematopoietic progenitor/stem cell lines that can generate erythroid, myeloid, and lymphoid cells upon transplantation into lethally irradiated mice. When transplanted into stem cell–deficient mice, these cell lines can generate a significant proportion of circulating erythrocytes in primary, secondary, and tertiary recipients for at least 18 months. Thus, Lhx2immortalizes multipotent hematopoietic progenitor/stem cells that can generate functional progeny following transplantation into lethally irradiated hosts and can long-term repopulate stem cell–deficient hosts.

Prostaglandin E2 Is a Potent Regulator of Vertebrate Hematopoietic Stem Cell Homeostasis.

Blood ◽  
2006 ◽  
Vol 108 (11) ◽  
pp. 680-680
Author(s):  
Trista E. North ◽  
Wolfram Goessling ◽  
Kamden R. Kopani ◽  
Claudia Lengerke ◽  
Carl R. Walkley ◽  
...  
Keyword(s):  
Stem Cells ◽  
Stem Cell ◽  
Hematopoietic Stem Cell ◽  
Embryonic Stem ◽  
Specific Effect ◽  
Biologically Active ◽  
Mead Acid ◽  
Cell Homeostasis ◽  
Hematopoietic Stem ◽  
Dose Dependent

Abstract Hematopoietic stem cells (HSCs) give rise to each of the blood lineages found in the adult vertebrate. Definitive HSCs are derived during embryogenesis in the aorta-gonad-mesonephros (AGM) region and subsequently colonize the niche in fetal and adult hematopoietic organs. To identify regulators of this process, a library of biologically active compounds was screened for effects on zebrafish HSC development. Embryos were exposed to individual chemicals (2498) from the 5-somite stage until 36 hpf, then processed by whole-mount in situ hybridization for the expression of HSC markers cmyb and runx1. Exposure to 47 of the compounds reduced the number of runx1+cmyb+ HSCs, while 35 increased this cell population. Several chemicals isolated in the screen were known to affect the prostaglandin (PG) synthesis pathway. Prostaglandin (PG) precursors, mead acid and linoleic acid, expanded HSCs, while fenbufen and celecoxib, inhibitors of cyclooxygenase (cox) activity, reduced HSC number. An examination of blood and vessel specific molecular markers demonstrated that angiogenesis was not altered significantly in treated embryos, illustrating the specific effect on HSCs. Exposure to exogenous PGE2 modestly increased HSCs, while use of a long-acting version of PGE2, 16,16-dimethylPGE2 (dmPGE2), markedly enhanced HSC number. Incubation of zebrafish embryos with known cox inhibitors confirmed the observation that inhibition of PG synthesis reduces HSCs. Injection of morpholino oligonucleotides designed against cox1, cox2 and PGE synthase inhibited the formation of HSCs in a dose dependent fashion, verifying that signaling through PGE2 is regulating AGM HSC number. Using CD41:GFP;LMO2:DsRed bigenic zebrafish, the modulation of HSCs following exposure to dmPGE2 or indomethacin was documented in vivo by confocal microscopy. Exposure to dmPGE2 following irradiation of adult fish induced HSC gene expression and enhanced hematopoietic recovery, indicating that PGE2 also functions in regulating adult marrow HSC homeostasis. Incubation of murine embryonic stem cells with PGE2 during embryoid body differentiation resulted in a dose-dependent increase of hematopoietic colonies in methylcellulose, while indomethacin exposure inhibited colony formation, demonstrating that the function of PGE2 in HSC regulation is conserved in mammals. Furthermore, ex vivo treatment of murine bone marrow with dmPGE2 resulted in a 2-fold increase in CFU-S number and enhanced the frequency of multi-lineage engrafting HSCs more than 3-fold in a limiting dilution competitive repopulation assay. The identification of the prostaglandin synthesis pathway as a potent regulator of vertebrate hematopoietic stem cell homeostasis has major implications for new treatments of diseases such as anemia and leukemia.

Embryonic and hematopoietic stem cells express a novel SH2–containing inositol 5′-phosphatase isoform that partners with the Grb2 adapter protein

Blood ◽  
2001 ◽  
Vol 98 (7) ◽  
pp. 2028-2038 ◽  
Author(s):  
Zheng Tu ◽  
John M. Ninos ◽  
Zhengyu Ma ◽  
Jia-Wang Wang ◽  
Maria P. Lemos ◽  
...  
Keyword(s):  
Stem Cells ◽  
Stem Cell ◽  
Embryonic Stem Cells ◽  
Hematopoietic Stem Cells ◽  
Hematopoietic Cells ◽  
Embryonic Stem ◽  
Cell Populations ◽  
Adapter Protein ◽  
Hematopoietic Stem ◽  
Stem Cell Populations

SH2–containing inositol 5′-phosphatase (SHIP) modulates the activation of immune cells after recruitment to the membrane by Shc and the cytoplasmic tails of receptors. A novel SHIP isoform of approximately 104 kd expressed in primitive stem cell populations (s-SHIP) is described. It was found that s-SHIP is expressed in totipotent embryonic stem cells to the exclusion of the 145-kd SHIP isoform expressed in differentiated hematopoietic cells. s-SHIP is also expressed in primitive hematopoietic stem cells, but not in lineage-committed hematopoietic cells. In embryonic stem cells, s-SHIP partners with the adapter protein Grb2 without tyrosine phosphorylation and is present constitutively at the cell membrane. It is postulated that s-SHIP modulates the activation threshold of primitive stem cell populations.

Clonal analysis of differentiating embryonic stem cells reveals a hematopoietic progenitor with primitive erythroid and adult lymphoid-myeloid potential

Development ◽  
2001 ◽  
Vol 128 (22) ◽  
pp. 4597-4604
Author(s):  
Rita C. R. Perlingeiro ◽  
Michael Kyba ◽  
George Q. Daley
Keyword(s):  
Stem Cells ◽  
Stem Cell ◽  
Hematopoietic Stem Cell ◽  
Embryoid Body ◽  
Embryonic Stem ◽  
Cell Types ◽  
Clonal Analysis ◽  
Erythroid Cell ◽  
Hematopoietic Stem

Embryonic stem (ES) cells differentiate into multiple hematopoietic lineages during embryoid body formation in vitro, but to date, an ES-derived hematopoietic stem cell has not been identified and subjected to clonal analysis in a manner comparable with hematopoietic stem cells from adult bone marrow. As the chronic myeloid leukemia-associated BCR/ABL oncogene endows the adult hematopoietic stem cell with clonal dominance without inhibiting pluripotent lymphoid and myeloid differentiation, we have used BCR/ABL as a tool to enable engraftment and clonal analysis. We show that embryoid body-derived hematopoietic progenitors expressing BCR/ABL maintain a primitive hematopoietic blast stage of differentiation and generate only primitive erythroid cell types in vitro. These cells can be cloned, and when injected into irradiated adult mice, they differentiate into multiple myeloid cell types as well as T and B lymphocytes. While the injected cells express embryonic (β-H1) globin, donor-derived erythroid cells in the recipient express only adult (β-major) globin, suggesting that these cells undergo globin gene switching and developmental maturation in vivo. These data demonstrate that an embryonic hematopoietic stem cell arises in vitro during ES cell differentiation that constitutes a common progenitor for embryonic erythroid and definitive lymphoid-myeloid hematopoiesis.

Regulated Expression of Activated Notch 1 during Embryonic Stem Cell Differentiation Preserves Multipotential Progenitors and Promotes Erythroid Cell Fate.

Blood ◽  
2004 ◽  
Vol 104 (11) ◽  
pp. 4151-4151
Author(s):  
Uma Ganapati ◽  
Lynne A. Bui ◽  
Maureen Lynch ◽  
Milana Dolezal ◽  
Hongying Tina Tan ◽  
...  
Keyword(s):  
Stem Cells ◽  
Stem Cell ◽  
Embryonic Stem Cells ◽  
Notch Signaling ◽  
Cell Fate ◽  
Embryonic Stem ◽  
Stem Cell Differentiation ◽  
Erythroid Cell ◽  
Hematopoietic Progenitors ◽  
Hematopoietic Stem

Abstract Hematopoietic stem cells pass sequentially through a series of developmental decision points regulating self-renewal and lineage-specific differentiation. In normal hematopoiesis proliferation is tightly linked to differentiation in ways that are poorly understood. The Notch gene family has been shown to be evolutionarily conserved and to play an important role in determining cell fate, survival, and proliferation in multiple organisms. Numerous in vitro and in vivo studies strongly support a role for Notch signaling in the regulation of stem cell signaling and hematopoiesis. To define the function of Notch in the earliest stages of hematopoiesis, a Tetracycline-inducible system regulating expression of a ligand-independent, constitutively active form of Notch1 was introduced into murine E14Tg2a embryonic stem cells. (Era and Witte, PNAS, 97;1737–1742,2000). During co-culture, OP9 stromal cells induce the embryonic stem cells to differentiate first to hemangioblasts and subsequently to hematopoietic cells. Our studies indicate that activation of Notch signaling in flk+ hemangioblasts dramatically reduces their proliferative capacity without inducing apoptosis. Furthermore, Notch1 activation significantly reduces the levels of hematopoietic stem cell markers CD34, c-Kit and the myeloid marker CD11b. These reversible effects suggest that Notch signaling maintains the hemangioblasts in an immature state and blocks hematopoietic differentiation. When activated Notch is induced in committed hematopoietic progenitors, a shift towards definitive erythroid differentiation and decreased myeloid differentiation is observed. Microarray analysis of day8 hematopoietic progenitors following Notch activation in hemangioblasts indicates upregulation of known downstream targets of Notch signaling. Based on these results, we propose that Notch signaling plays a critical role in the earliest events regulating hematopoiesis.

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