Elaine Dzierzak

Characterization of the first definitive hematopoietic stem cells in the AGM and liver of the mouse embryo.

At day 10 in mouse gestation, the intraembryonic aorta-gonads-mesonephros (AGM) region generates the first definitive hematopoietic stem cells (HSCs) of the adult blood system. By 11 days postcoitum, the liver contains such HSCs. While HSCs of the adult bone marrow and late-stage fetal liver have been extensively characterized for cell surface markers, there has been no phenotypic description of the first HSCs during embryo development. We report here the temporal cell surface phenotype of HSCs from the AGM region and early fetal liver and show that all HSCs reside in the c-kit+ population. c-kit+ HSCs from AGM and liver are mainly CD34+ and in the AGM are in both Mac-1+ and Mac-1 fractions. These results demonstrate that during mouse ontogeny the first definitive HSCs are similar in cell surface phenotype to the HSCs of adult bone marrow but that spatial localization and developmental time are critical factors in the phenotypic assessment of this functional cell population.

Mouse embryonic hematopoiesis

The hematopoietic system of vertebrates is derived from the mesodermal germ layer in early embryogenesis. Various animal models have been used for the study of hematopoiesis, from early stages in the visceral yolk sac or its analog, to the later stages where hematopoiesis is observed in intraembryonic areas surrounding the aorta, genital ridge and pro/mesonephros. Using the mouse as a model, we describe what is known about mammalian embryonic hematopoiesis and put it in the context of hematopoietic cell formation in avian, amphibian and fish embryos. Evolutionary comparisons and recent experimental evidence show that there are two embryonic sites of developing hematopoietic activity in the mouse before fetal liver hematopoiesis and suggest that, during ontogeny, two successive waves of hematopoietic activity may contribute to the blood system of the adult.

Human Placenta Is a Potent Hematopoietic Niche Containing Hematopoietic Stem and Progenitor Cells throughout Development

Hematopoietic stem cells (HSCs) are responsible for the life-long production of the blood system and are pivotal cells in hematologic transplantation therapies. During mouse and human development, the first HSCs are produced in the aorta-gonad-mesonephros region. Subsequent to this emergence, HSCs are found in other anatomical sites of the mouse conceptus. While the mouse placenta contains abundant HSCs at midgestation, little is known concerning whether HSCs or hematopoietic progenitors are present and supported in the human placenta during development. In this study we show, over a range of developmental times including term, that the human placenta contains hematopoietic progenitors and HSCs. Moreover, stromal cell lines generated from human placenta at several developmental time points are pericyte-like cells and support human hematopoiesis. Immunostaining of placenta sections during development localizes hematopoietic cells in close contact with pericytes/perivascular cells. Thus, the human placenta is a potent hematopoietic niche throughout development.

Impaired embryonic haematopoiesis yet normal arterial development in the absence of the Notch ligand Jagged1.

Specific deletion of Notch1 and RBPjκ in the mouse results in abrogation of definitive haematopoiesis concomitant with the loss of arterial identity at embryonic stage. As prior arterial determination is likely to be required for the generation of embryonic haematopoiesis, it is difficult to establish the specific haematopoietic role of Notch in these mutants. By analysing different Notch‐ligand‐null embryos, we now show that Jagged1 is not required for the establishment of the arterial fate but it is required for the correct execution of the definitive haematopoietic programme, including expression of GATA2 in the dorsal aorta. Moreover, successful haematopoietic rescue of the Jagged1‐null AGM cells was obtained by culturing them with Jagged1‐expressing stromal cells or by lentiviral‐mediated transduction of the GATA2 gene. Taken together, our results indicate that Jagged1‐mediated activation of Notch1 is responsible for regulating GATA2 expression in the AGM, which in turn is essential for definitive haematopoiesis in the mouse.

Hematopoietic stem cells and their precursors: developmental diversity and lineage relationships

Summary:  Within the context of the developing embryo, restrictions in cell lineage potential occur through cell–cell interactions and signaling molecules, leading to changes in genetic programs and to the emergence of disparate tissues containing functionally distinct cell types including somatic stem cells. Tissue maintenance in the adult is thought to occur through specific stem cells, and in the case of the hematopoietic system, through hematopoietic stem cells (HSCs). These cells arise in midgestation within the region of the embryo containing the dorsal aorta, gonads, and mesonephros (AGM) and are thought to maintain a distinct hematopoietic lineage‐restricted fate. However, recent transplantation experiments suggest that within the adult, HSCs previously thought to be restricted can, under certain circumstances, display unexpected lineage potentials. With these surprising and controversial results, it is becoming apparent that a better understanding of the developmental processes, molecular programs and lineage relationships leading to the emergence of adult stem cells will provide insight into the incremental steps involved in lineage determination, and perhaps possibilities for the manipulated differentiation of stem cells. The most widely studied, accessible stem cell and cellular differentiation hierarchy is that of the hematopoietic system. With the issue of stem cell potential in the forefront, the focus of this review is on the development of the hematopoietic system: how HSCs arise in the embryo, the lineage relationships of hematopoietic cells as they are generated, and the identification of precursor cells fated to the hematopoietic lineage throughout ontogeny.

The emergence of definitive hematopoietic stem cells in the mammal.

Purpose of reviewHematopoietic stem cells (HSC) are the basis for blood formation during adult life. The amazing potency of HSCs has been exploited for over 30 years in regenerative therapies for patients with blood-related genetic disease and leukemia. As clinically important cells and also as the most widely studied cell differentiation system, they have been the focus of intense fundamental research. Indeed, HSC research has established many paradigms in the more general field of stem cells. Recently, the study of the embryonic origins of HSCs and their genetic program is beginning to provide unique insights into how these stem cells are formed, maintained, and expanded, and how they contribute to the complex adult hematopoietic system. Although many short-lived hematopoietic progenitors are present in early stage mammalian embryos, this review will focus on the events leading to emergence of the most potent cells of the hematopoietic system, HSCs and on their developmental lineage relationships. Recent findingsDevelopmental and genetic studies further our understanding of the fate determination events occurring in several embryonic tissues leading to the generation of potent HSCs - those cells with the ability to long-term, high-level repopulate all hematopoietic lineages of the adult. SummarySeveral mammalian embryonic tissues contribute to the growth and/or generation of potent HSCs that are the source of blood cells throughout the lifespan of the individual. Insight into how mammalian HSC fate is determined has been provided through functional, phenotypic, and genetic studies at early developmental stages.

The EMT regulator Zeb2/Sip1 is essential for murine embryonic hematopoietic stem/progenitor cell differentiation and mobilization

Zeb2 (Sip1/Zfhx1b) is a member of the zinc-finger E-box-binding (ZEB) family of transcriptional repressors previously demonstrated to regulate epithelial-to-mesenchymal transition (EMT) processes during embryogenesis and tumor progression. We found high Zeb2 mRNA expression levels in HSCs and hematopoietic progenitor cells (HPCs), and examined Zeb2 function in hematopoiesis through a conditional deletion approach using the Tie2-Cre and Vav-iCre recombination mouse lines. Detailed cellular analysis demonstrated that Zeb2 is dispensable for hematopoietic cluster and HSC formation in the aorta-gonadomesonephros region of the embryo, but is essential for normal HSC/HPC differentiation. In addition, Zeb2-deficient HSCs/HPCs fail to properly colonize the fetal liver and/or bone marrow and show enhanced adhesive properties associated with increased β1 integrin and Cxcr4 expression. Moreover, deletion of Zeb2 resulted in embryonic (Tie2-Cre) and perinatal (Vav-icre) lethality due to severe cephalic hemorrhaging and decreased levels of angiopoietin-1 and, subsequently, improper pericyte coverage of the cephalic vasculature. These results reveal essential roles for Zeb2 in embryonic hematopoiesis and are suggestive of a role for Zeb2 in hematopoietic-related pathologies in the adult.

Whole-transcriptome analysis of endothelial to hematopoietic stem cell transition reveals a requirement for Gpr56 in HSC generation.

Using highly sensitive RNAseq to examine the whole transcriptome of enriched aortic hematopoietic stem cells and endothelial cells, the authors find G-protein–coupled receptor, Gpr56, is required to generate the first HSCs during endothelial to hematopoietic cell transition.

Ontogenic emergence of definitive hematopoietic stem cells

Research in the past 10 years has dramatically increased our knowledge of the development of the mammalian hematopoietic system and has provided insight into the embryonic sites of hematopoietic cell generation, the variety of hematopoietic cell types produced, and some of the microenvironmental influences on the rapidly growing blood system. Indeed, within mammalian embryos, it is now widely accepted that the embryo proper produces the first adult repopulating hematopoietic stem cells. This mesodermally derived intraembryonic region, known as the aorta-gonad-mesonephros region or, at a slightly earlier developmental stage, the paraaortic splanchnopleura, produces, respectively, potent hematopoietic stem cells and multipotent progenitor cells before their appearance in the yolk sac. This review focuses on the most recent findings concerning qualitative and quantitative aspects of hematopoietic stem-cell development, the endothelium as a possible direct precursor population of hematopoietic stem cells, and the microenvironment leading to the onset and maintenance of hematopoietic stem cells in the mammalian embryo.

Embryonic Beginnings of Definitive Hematopoietic Stem Cells

Abstract: The ability of the many cell types within the adult blood system to be constantly replenished and renewed from hematopoietic stem cells is an interesting problem in development and differentiation and has led to questions concerning how, when and where thses stem cells for the adult hematopoietic system are generated within the embryo. During embryonic development many mature hematopoietic cells appear before adult‐type hematopoietic stem cells thus the notion of a conventional hematopoietic hierarchy is challegned. Experiments probing the development of hematopoietic stem cells in the mouse embryo strongly suggest that at least two independent hematopoietic sites generate blood cells during development; the yolk sac, which produces the transient embryonic hematopoietic system, and the AGM (aorta‐gonad‐mesonephros) region, which initiates the long‐lived adult hematopoietic system.