|
|||||||||
|
|
|
|
|
|
|
|
|
|
|
Prepublished online as a Blood First Edition Paper on September 19, 2002; DOI 10.1182/blood-2002-06-1699.
HEMATOPOIESIS
From the Department of Hematology/Oncology, Children's
Hospital; Department of Pediatric Oncology, Dana Farber Cancer
Institute; and Howard Hughes Medical Institute, Boston, MA.
Murine hematopoietic stem cells (HSCs) originate from
mesoderm in a process that requires the transcription factor SCL/Tal1. To define steps in the commitment to blood cell fate, we compared wild-type and SCL Blood cell development during embryogenesis is
initiated within the mesodermal germ layer in a process that requires
the basic helix-loop-helix (bHLH) transcription factor
SCL/tal1.1-3 However, the precise steps leading to
commitment to blood cell fate are incompletely characterized. Intimate
spatial and temporal development of the hematopoietic and endothelial
cell lineages in the embryo, shared marker gene expression, and
knockout mice with defects in both lineages have led to the hypothesis
of a common origin for blood and endothelium, the
hemangioblast.4-12 By definition, the hemangioblast is a
common progenitor for primitive and definitive hematopoietic and
endothelial cell lineages. The divergence of the lineages and the
phenotype of the earliest precursors in each lineage, however, are not
fully defined.
In the mouse embryo, the commitment to hematopoiesis occurs within days
7 to 8.25 (E7-E8.25) of embryonic development, during which time
hematopoietic progenitors of primitive and definitive hematopoietic
lineage appear in the extraembryonic yolk sac and the embryo
proper.13,14 After a transient wave of primitive erythropoiesis in the yolk sac, definitive hematopoietic progenitors from the yolk sac and para-aortic
splanchnopleural/aorta-gonad-mesonephros (PAS/AGM) region seed the
fetal liver, which is the principal site of definitive hematopoiesis
during fetal life.15-17 During the fetal liver stage,
hematopoietic stem cells acquire the capacity to engraft and
reconstitute bone marrow (BM) and thus support lifelong hematopoiesis.
The origin of hematopoietic stem cells (HSCs) during development is a
subject of controversy. It has been generally accepted that de novo
hematopoiesis occurs in 2 independent sites, the extraembryonic yolk
sac and the intraembryonic PAS/AGM and that cells originating from both
locations can contribute to adult hematopoiesis when stimulated
appropriately. The earliest site of detectable HSC activity in
developing mouse embryo is the PAS/AGM region, where adult
reconstituting HSCs can be found at E10.5, before the fetal liver
stage.17-21
Although no adult BM reconstituting stem cells are found in the
yolk sac before fetal liver hematopoiesis, yolk sac cells can
reconstitute multilineage, long-term hematopoiesis in conditioned newborn recipients.22,23 This has led to the hypothesis
that yolk sac stem cells are immature and require an additional
maturation step within fetal liver before BM engraftment can be
achieved. Yolk sac cells have been shown to acquire some adult BM
reconstitution potential by coculture on an AGM-derived stromal cell
line24 or by overexpression of HOXB4.25
In recent years, model systems have been developed that permit
the study of hematopoietic and endothelial commitment in
vitro.26-32 Murine embryonic stem (ES) cells grown in
suspension culture without feeder cells and leukemia inhibitory factor
(LIF) spontaneously form cell aggregates called embryoid bodies (EBs)
and differentiate into cell types representing all germ layers,
including hematopoietic and endothelial cells. The EBs form cystic
structures resembling yolk sac blood islands where the first steps of
hematopoietic development occur in a fashion similar to that in the
mouse embryo. Furthermore, in vitro differentiation of ES cells affords
study of gene-targeted cells without confounding embryonic lethality of
the developing mouse. In spite of the close similarity of hematopoietic development in the EBs to hematopoiesis in vivo, the validity of this
model system has been questioned because of a failure to demonstrate
the presence of authentic hematopoietic stem cells. Recent work shows
that EB-derived hematopoietic progenitors can achieve some adult BM
reconstitution potential by transient overexpression of HOXB4 or
BCR-ABL.25,33
Studies in gene-targeted mice and ES cells have demonstrated a critical
role for the bHLH transcription factor SCL in the initiation of the
blood program. Without SCL, no blood cells of any lineage develop in
vivo in chimeric mice or in vitro in embryoid bodies.34,35
In contrast, endothelial cells are present but fail to remodel properly
in the yolk sac.36 In vitro study of SCL Culture and differentiation of ES cells
FACS analysis and sorting
Methylcellulose colony assays EBs were harvested at specific time points for plating into the blast cell colony assay (days in embryoid body cultures in vitro [d]; d3.25-3.5) or hematopoietic colony assays (d5-7). For blast cell colony assay, the cells were plated at 0.75 to 1.5 × 105 cells/mL in 1% methylcellulose containing 10% FCS (Summit Biotechnologies), 4.5 × 10 4
M MTG, 1% penicillin/streptomycin, 1% L-glutamine supplemented with 5 ng/mL vascular endothelial growth factor (VEGF), 5 ng/mL interleukin-6 (IL-6), 100 ng/mL stem cell factor (SCF; R&D Systems, Minneapolis, MN), 25 µg/mL ascorbic acid, and 300 µg/mL transferrin (Roche) to support hemangioblast differentiation. To verify
hematopoietic and endothelial cell potential in the blast cell
colonies, individual blast colonies were picked at 4 days of
methylcellulose culture and were plated into growth-factor-reduced
Matrigel (Becton Dickinson)-coated dishes in IMDM, 10% FCS, 10%
horse serum, 5 ng/mL VEGF, 10 ng/mL insulinlike growth factor-1
(IGF-1), 10 ng/mL basic fibroblast growth factor (bFGF), 2 U/mL
erythropoietin (EPO), 50 ng/mL SCF, 10 ng/mL IL-3 (R&D Systems), and
endothelial cell growth supplement (Collaborative Biomedical Products,
Becton Dickinson).
For analysis of hematopoietic precursors, the methylcellulose mix was supplemented with hematopoietic growth factors 2 U/mL EPO, 100 ng/mL mSCF, 1 ng/mL mIL-3, 5 ng/mL murine IL-6 (mIL-6), 5 ng/mL murine thrombopoietin (mTPO), 30 ng/mL human granulocyte-colony-stimulating factor (hG-CSF), 3 ng/mL murine granulocyte macrophage-colony-stimulating factor (mGM-CSF), 5 ng/mL IL-11, and 5 ng/mL mM-CSF. Hematopoietic colonies were scored individually by microscopy and May-Grünwald-Giemsa staining at 5 to 8 days of differentiation. Endothelial cell cultures FACS-sorted cells from EBs were plated on Matrigel-coated dishes (Becton Dickinson) in DMEM, 10% FCS (Summit), penicillin/streptomycin, 25 ng/mL VEGF, 10 ng/mL IGF, 10 ng/mL bFGF (R&D Systems), and endothelial cell growth supplement (Collaborative Biomedical Products, Becton Dickinson). Endothelial cell potential was monitored by the growth of adherent cells and FACS staining for Flk1 and PECAM (CD31) (Becton Dickinson).Analysis of mouse embryos To analyze embryonic and fetal hematopoietic development in mice, inbred (C57Bl6) and outbred (Swiss Webster) mice were time-mated, and embryos were collected at E8.5, E9.5, and E10.5 for studies of yolk sac hematopoiesis and at E12.5 or E14.5 for fetal liver hematopoiesis. Yolk sacs were dissected from the embryo proper, trypsinized, separated into single-cell suspension by a 21-gauge syringe, filtered, and analyzed by FACS. Fetal livers were collected and separated into single-cell suspension by pipetting and filtration.Wholemount immunohistochemistry for E8.5 and E9.5 embryos using anti-CD41 antibody (PharMingen) was performed essentially as described.36 Peroxidase staining was used for antibody detection. Fluorescein isothiocyanate (FITC)-conjugated anti-CD41 antibody was used to stain 10-µm cryosections of dissected E9.5 yolk sac that had been fixed with 4% paraformaldehyde.
Kinetics of expression of hematopoietic and endothelial surface markers in wild-type and transcription factor mutant EBs To analyze the commitment of mesoderm to hematopoietic and endothelial cell lineages, wild-type and transcription factor mutant ES cells were examined during differentiation in vitro. SCL/ ES cells were used as a negative control for
hematopoietic development because previous studies have shown complete
absence of hematopoietic activity in SCL/
EBs.3,34,35 EBs were harvested at specific times and
subjected to FACS analysis. Table 1
demonstrates similar kinetics of expression of the early
mesoderm/hemangioblast markers c-kit, Flk1, and  4-integrin (CD49d)
in wild-type and SCL/ EBs. Similarly, endothelial cell
markers endoglin (CD105), MECA32, and VE-cadherin and combined
endothelial/hematopoietic markers CD34 and PECAM (CD31) were expressed
in wild-type and SCL/ EBs. However, all markers
specific for hematopoietic cells (CD41, CD45, and Ter119) were absent
in SCL/ EBs. In these temporal studies, CD41 was
identified as the earliest blood-specific marker in the wild-type EBs.
Surface expression of CD41 was first detected at approximately d4.25 to
d4.75 of EB differentiation and increased progressively to 5% to 30%
by d6 to d7 (Table 1; Figure 1).
RT-PCR analysis for CD41 demonstrated a correlation of CD41
surface expression with the presence of CD41 mRNA in the EBs. In
wild-type EBs, the expression of CD41 RNA was detected approximately 12 hours after the appearance of SCL RNA transcripts, whereas no CD41 RNA
was found in SCL Combined analysis of CD41 and other hematopoietic and endothelial
markers showed that CD41+ cells developed from
c-kit+ cells in the EBs with partial overlap with Flk1
expression. All CD41+ cells coexpressed We also examined CD41 expression during in vitro differentiation of
knockout ES cells for runx1/AML1.39 Whereas no expression of CD41 was detected in SCL Characterization of hematopoietic and endothelial surface marker expression in blast cell colonies EB-derived blast cell colonies that represent early SCL-dependent hemangioblast/hematopoietic activity in cultures with VEGF were analyzed for surface expression of hematopoietic and endothelial cell markers. Cells from d3.25 to d3.5 EBs were first cultured in methylcellulose with VEGF, IL-6, and SCF. Four days later, cultures were scored for the presence of colonies containing loosely attached cells characteristic of blast cell colonies. Hematopoietic and endothelial potential in the blast cell colonies was verified by plating individual colonies into Matrigel with hematopoietic and endothelial growth factors as previously described.32 As shown earlier,3,37 SCL/ EBs lack the
potential to form blast cell colonies. The phenotype of the cells in
blast colonies was analyzed by pooling the loosely attached cells from
methylcellulose cultures and subjecting them to FACS analysis. Although
the cells that give rise to blast colonies are Flk1
positive37 (data not shown), only a portion of the nonadherent cells in their progeny (40% ± 4%) express Flk1,
demonstrating down-regulation of Flk1 during hematopoietic development.
In comparison, 4-integrin, PECAM, and CD41 were up-regulated during
blast colony development and were expressed on the surfaces of most
(more than 85%) cells (Figure 2). Other
hematopoietic markers were present in lower frequencies (Mac1
42% ± 3%, Gr1 41% ± 5%, Ter119 6% ± 1.4%, CD45
23% ± 3%, c-kit 25% ± 9%). A subfraction of cells in the
blast colonies also expressed the endothelial markers MECA32 (58% ± 6%) and VE-cadherin (42% ± 19%) and the
endothelial/hematopoietic marker CD34 (34% ± 5%) (Figure
2C).
Developmental potential of CD41+ and CD41 /
EBs and the most frequently expressed blood-specific marker in
SCL-dependent blast colonies, wild-type EBs were fractionated based on
CD41 expression, and the developmental potentials of CD41+
and CD41 populations were individually assessed. EBs were
differentiated for 5 to 7 days, trypsinized into single-cell
suspensions, and sorted for surface expression of CD41. Expression of
CD41 was present on 2% to 4% of cells at day 5 and increased to 5%
to 30% at days 6 and 7. Morphologically, EB-derived CD41+
cells comprised a homogenous, blastlike population (Figure
3A). In contrast, CD41
cells were heterogeneous in appearance (Figure 3B).
To assay hematopoietic potential, sorted cells were plated into
methylcellulose cultures supplemented with a combination of hematopoietic growth factors. At all times, definitive hematopoietic activity was highly enriched in the CD41+ fraction (Figure
3C), whereas the CD41 To study the potential of CD41+ cells to differentiate into
endothelial cells, CD41+ and CD41 Expression of CD41 in the mouse embryo To correlate in vitro ES cell differentiation with embryonic hematopoiesis in vivo, CD41 expression was studied in time-mated mouse embryos. Wholemount staining demonstrated the expression of CD41 in yolk sac blood islands at E8.5 and E9.5 (not shown and Figure 4) in locations similar to that of GATA-1 (not shown). At E9.5, CD41 expression was detected in the vitelline and umbilical vessels; however, the endothelial cell network that stains with PECAM (not shown) in the embryo proper did not express detectable CD41. Staining with FITC-conjugated anti-CD41 antibody demonstrates the presence of individual CD41+ cells inside yolk sac blood islands at E9.5 (Figure 4B) and occasionally inside the vessels in the embryo proper (not shown). Endothelial cells in blood islands at E9.5 did not express detectable CD41.
FACS analysis of hematopoietic organs revealed abundant
(5%-25%) surface expression of CD41 in the yolk sac at E8.5, E9.5, and E10.5. Similarly, some CD41 expression was detectable in the AGM
region at E10.5, and in E12.5, and E14.5 fetal liver, indicating localization of CD41 expression into sites of hematopoietic activity in
the mouse embryo and the fetus (Figure 5A
and not shown).
Definition of the surface phenotype of hematopoietic progenitors during murine ontogeny Analysis of the kinetics of surface marker expression in the EBs and yolk sac demonstrated that CD45 expression is first detectable in a subpopulation of CD41+ cells (Figures 1C, 5A). To define whether hematopoietic progenitor activity resided only in the CD45+ fraction of CD41+ cells, cells were sorted by CD45 and CD41 expression. Surprisingly, in E6 EBs and E9.5 yolk sac, clonogenic progenitor activity was detected also in CD45 cells (76% vs 22% in CD45
CD41+ and CD45+CD41+ subfractions
in EBs, and 33% vs 62% in the yolk sac, respectively; Figure 5B).
These results indicate that the first hematopoietic progenitors during
ontogeny did not express CD45, a panhematopoietic marker in the adult.
In contrast, in E14.5 fetal liver, all colony-forming activity resided
in the CD45+ fraction, with only a subpopulation
coexpressing CD41 (66% in CD45+CD41 vs 32%
in CD45+CD41+ fractions). In EBs and yolk sac
(YS), CD45+CD41+ and CD45
CD41+ fractions gave rise to multilineage colonies with
erythroid, megakaryocytic, and myeloid elements (not shown and Figure
5C-D). In contrast, in fetal liver, the CD41+ fraction
rarely gave rise to multilineage colonies but mainly consisted of
myeloid progenitors that differentiated into mast cells and macrophages
in culture, whereas multilineage CFU-C were enriched in the
CD45+CD41 fraction (not shown).
Multicolor FACS of EBs and yolk sac cells demonstrated the coexpression
of stem cell and progenitor cell markers c-kit and CD34 with CD41 (not
shown and Figure 6). Fractionation of the CD41+ population by positive c-kit expression increased the
CFU-C frequency and showed that most definitive hematopoietic
progenitors in the EBs and yolk sac are included within the
CD41+ c-kit+ population (Table
2).
SCL/tal 1 is strictly required for expression of CD41, whereas runx1/AML1 is essential for development of CD41+ c-kit+ definitive hematopoietic progenitors Studies of gene-targeted mice have proved pivotal in defining the stages of commitment toward hematopoiesis and within the hematopoietic system. Such analyses have identified major regulatory molecules that direct blood cell development.40 SCL/Tal1, a bHLH transcription factor whose expression is deregulated by chromosomal translocations in T-cell leukemia,41 is essential for initiation of the hematopoietic program at the putative hemangioblast stage.3 In comparison, the absence of runx1/AML1, also known as core-binding factor (CBF) A2, leads to the loss of definitive hematopoietic lineages, whereas primitive erythroid development is relatively unaffected.39,42Our aim here was to study events downstream of SCL and AML1 to identify
the phenotype of the first hematopoietic progenitors arising during
development. On differentiation of wild-type and SCL-deficient embryoid
bodies, we identified CD41 (GpIIb) as the earliest surface marker
missing from SCL Comparison of CD41 expression in other transcription factor mutants with blocks in various stages of hematopoietic development demonstrates the sustained expression of CD41 in the absence of runx1/AML1, GATA-1, and GATA-2 (Figure 1; Y.F. and S.H.O., unpublished data, January 2001). The failure of runx1/AML1-deficient EBs to develop CD41+c-kit+ double-positive cells suggests that CD41 and c-kit mark definitive hematopoietic progenitors in the embryo. This hypothesis is supported by progenitor assays of sorted subpopulations in which definitive hematopoietic colonies in the d6 EBs and E9.5 yolk sac are highly enriched in the CD41+c-kit+ double-positive fraction. Expression of CD41 appears after the hemangioblast stage and development of primitive hematopoietic and endothelial cell lineages The earliest hematopoietic activity detected in vitro from the EBs is derived from Flk1+ cells, which, in response to VEGF, give rise to blast cell colonies that exhibit definitive and primitive hematopoietic and endothelial cell potential, filling the formal criteria for the hemangioblast. Because most of the daughter cells in blast cell colonies also express CD41, we considered the possibility that CD41 might be a marker for the hemangioblast. However, expression of CD41 in the EBs appears after the detection of hemangioblast and primitive erythroid activity (d3.25-d3.5 and d4, respectively)31 and endothelial marker expression. Although early CD41+ cells from the EBs occasionally gave rise to primitive erythroid colonies, as did CD41 cells,
our data do not support a consistent expression pattern for CD41 with
respect to primitive erythroid development in the embryoid bodies. In
contrast, the kinetics and abundant expression of CD41 in the EBs
correlated with the development of definitive hematopoietic
progenitors. Likewise, in the yolk sac, the appearance of CD41 is
delayed from the peak of primitive erythroid progenitor activity
(E7-E8).13 Furthermore, FACS analysis with CD41 and Ter119
in the yolk sac demonstrate distinct expression patterns of these
markers, suggesting that maturing primitive erythroid cells do not
express CD41.
The endothelial cell markers endoglin, PECAM, and MECA32 appear in EBs
approximately 1 day earlier than CD41, suggesting that the endothelial
cell lineage develops before the development of CD41+
cells. Furthermore, adherent cells expressing Flk1 and PECAM developed
from the CD41 All CD41+ cells in EBs and blast colonies also expressed
CD41+ cells in EBs and yolk sac coexpress markers of definitive hematopoiesis (CD45) and neonatal reconstituting stem cells (c-kit and CD34) In adult hematopoiesis, CD45 is generally considered a panhematopoietic marker. Our data show that in the EBs and in the yolk sac, the expression of CD45 appears exclusively in CD41+ cells approximately 1 to 2 days later than CD41. Interestingly, in EBs and in the yolk sac, a significant proportion of colony-forming activity was detected in the CD45 CD41+
population. Furthermore, the expression of CD45 did not correlate with
a gain in differentiation potential in culture because
CD45+ and CD45 fractions gave rise to mixed
colonies and to single-lineage definitive erythroid, megakaryocyte, and
myeloid colonies. These data suggest that in vitro in EBs and in vivo
in the yolk sac, the expression of CD41 spans the development from a
Flk1+ hemangioblast to definitive hematopoietic progenitors
that later express CD45 (Figure 7). One
obvious caveat is that our data are derived from progenitor assays,
whereas the development and surface phenotype of HSCs remains
to be studied by in vivo assays. Nevertheless, newborn reconstituting
stem cells within the yolk sac have been shown to express c-kit and
CD34,23 markers that we found to be coexpressed with CD41
in the yolk sac and in EBs. During mouse development, it has been
proposed that the earliest adult repopulating HSCs arise from the walls
of dorsal aorta in the AGM region and vitelline and umbilical
vessels.17-21 Wholemount staining and FACS analyses showed
some expression of CD41 in the vitelline and umbilical vessels and in
the AGM region. In situ immunohistochemistry analysis of CD41
expression during mouse embryonic development demonstrates that CD41 is
expressed in hematopoietic clusters in the walls of dorsal aorta and
umbilical artery and in the lumen of yolk sac blood
islands.44 The developmental potential of the early CD41+ cells in the mouse remains to be addressed by in vivo
assay. Interestingly, North et al45 recently reported the
presence of CD45 HSCs in E10.5 AGM and E12.5 fetal liver,
a phenomenon that was accentuated by runx1/AML1 haploinsufficiency.
Whether these early HSCs express CD41 remains to be studied
further.
Expression of CD41 is down-regulated in hematopoietic progenitors by the fetal liver stage The fetal liver serves as the site of definitive hematopoietic development and HSC maturation from E11.5 to early postnatal life. Abundant adult reconstituting HSCs are found in the fetal liver from E12.517,20,46 We extended the sorting analysis with CD45 and CD41 expression to E14.5 fetal liver and performed colony-forming unit assays from sorted subpopulations. In contrast to embryoid bodies and yolk sac in which a significant fraction of hematopoietic progenitors do not express CD45, all hematopoietic activity was detected within the CD45+ population in the E14.5 fetal liver. When subfractionated with CD41 expression, most CFU-C, including mixed colonies and CFU-S8, were found in the CD45 single-positive population (H.K.A.M. and S.H.O., unpublished data, August 2001). Further long-term transplantation studies are required to determine whether CD41 is expressed in adult BM reconstituting stem cells in the fetal liver.In humans, CD41 cells in cord blood exhibit myeloid and lymphoid potential long-term culture initiating cell (LTC-IC) and nonobese diabetic/severe combined immunodeficient (NOD-SCID) repopulating activity.47 In the avian model, CD41+CD45+ cells also develop from intra-aortic clusters, the sites of HSC emergence, and in embryonic and adult BM exhibit multilineage potential.48 These data suggest that across species, CD41 expression is not restricted to the megakaryocyte lineage. At the time of initiation of definitive hematopoiesis, CD41 is expressed in most definitive hematopoietic progenitors, whereas later in development its expression is restricted to a subset of progenitors with more restricted developmental potential.47-49 Differential expression of hematopoietic surface markers during ontogeny has been described for other surface markers, such as CD34, Mac1, and AA4.20,50,51 The functional significance of such variation in relation to specific microenvironment influences has yet to be fully explored, though CD34 positivity appears to correlate with stem cell activation.52 Thus far, no markers that are predominantly associated with the yolk sac stage have been identified. Whether down-regulation of CD41 is involved in the maturation of adult BM reconstituting stem cells remains to be studied by further in vivo assays. Role of CD41 in hematopoiesis during ontogeny is unknown We have identified CD41 as a commitment marker for hematopoiesis between Flk1+ hemangioblasts and CD45+ hematopoietic cells during mouse ontogeny (Figure 7). Sorting for CD41 and c-kit enables the isolation of cell populations from EBs or yolk sac in which early hematopoietic activity can be studied. Further in vivo studies are required to determine whether CD41+ hematopoietic cells represent a transient progenitor population during development or whether they serve as precursors for the developing stem cells. A relationship between CD41 expression and the establishment of definitive hematopoiesis is supported by gene-targeting studies. Expression of Cre recombinase under the control of regulatory elements of the CD41 gene results in blood-specific recombination between loxP sites and the expression of a Cre-activatable reporter gene in all lymphoid and myeloid cells in the fetus.53 If CD41+ cells are the earliest precursors for developing HSCs in the mouse embryo and during in vitro differentiation of mouse ES cells, evaluation of the expression of CD41 during hematopoietic development in human embryos and ES cells will be of interest. In support of the relevance of CD41 as a marker for developmental hematopoiesis in humans, CD41+ cells from human cord blood have been shown to exhibit reconstitution potential.47 Furthermore, in the era of stem cell plasticity and transdifferentiation studies, the possibility of existence of CD45CD41+ hematopoietic progenitors/stem
cells during the transdifferentiation process should be considered.
Besides the usefulness of CD41 as a marker of the definitive
hematopoietic lineage in the embryo, the functional role of CD41 during
hematopoietic development is still unknown. If CD41 is expressed and
functionally important in early embryonic hematopoiesis, it is perhaps
surprising that CD41 knockout mice fail to demonstrate any striking
abnormalities in hematopoiesis other than a platelet function
defect.54 This suggests that lack of CD41 and signaling through the CD41CD61 (GPIIbIIIa) complex are not selected against during hematopoietic development. However, it is possible that in
hematopoietic progenitors, the function of CD41 can be partially replaced by an alternative dimerization partner for CD61. Recent studies with mast cells in CD41 knockout mice show that the absence of
CD41 in BM mast cells resulted in a compensatory increase in
S.H.O. is an Investigator of the Howard Hughes Medical Institute.
We thank Gordon Keller and Marion Kennedy for help in setting up the
blast cell colony assay. We also thank Nancy Speck and Gordon Keller
for the runx1/AML
Submitted June 6, 2002; accepted August 14, 2002.
Prepublished online as Blood First Edition Paper, September 19, 2002; DOI 10.1182/blood-2002-06-1699.
The publication costs of this article were defrayed in part by page charge payment. Therefore, and solely to indicate this fact, this article is hereby marked "advertisement" in accordance with 18 U.S.C. section 1734.
Reprints: Stuart H. Orkin, Department of Pediatric Oncology, Dana-Farber Cancer Institute, 44 Binney St, Boston, MA, 02115; e-mail: stuart_orkin{at}dfci.harvard.edu.
1. Shivdasani RA, Mayer EL, Orkin SH. Absence of blood formation in mice lacking the T cell leukaemia oncoprotein tal-1/SCL. Nature. 1995;373:432-434[CrossRef][Medline] [Order article via Infotrieve].
2.
Robb L, Lyons I, Li R, et al.
Absence of yolk sac hematopoiesis from mice with a targeted disruption of the scl gene.
Proc Natl Acad Sci U S A.
1995;92:7075-7079 3. Robertson SM, Kennedy M, Shannon JM, Keller G. A transitional stage in the commitment of mesoderm to hematopoiesis requiring the transcription factor SCL/tal-1. Development. 2000;127:2447-2459[Abstract]. 4. Sabin FR. Studies of the origin of blood vessels and of red corpuscles as seen in the living blastoderm of the chick during second day of incubation. Contrib Embryol. 1920;9:213-262.
5.
Pardanaud L, Yassine F, Dieterlen-Lievre F.
Relationship between vasculogenesis, angiogenesis and haemopoiesis during avian ontogeny.
Development.
1989;105:473-485 6. Haar JL, Ackerman GA. A phase and electron microscopic study of vasculogenesis and erythropoiesis in the yolk sac of the mouse. Anat Rec. 1971;170:199-223[CrossRef][Medline] [Order article via Infotrieve].
7.
Young PE, Baumhueter S, Lasky LA.
The sialomucin CD34 is expressed on hematopoietic cells and blood vessels during murine development.
Blood.
1995;85:96-105
8.
Eichmann A, Corbel C, Nataf V, Vaigot P, Breant C, Le Douarin NM.
Ligand-dependent development of the endothelial and hemopoietic lineages from embryonic mesodermal cells expressing vascular endothelial growth factor receptor 2.
Proc Natl Acad Sci U S A.
1997;94:5141-5146 9. Takakura N, Huang XL, Naruse T, et al. Critical role of the TIE2 endothelial cell receptor in the development of definitive hematopoiesis. Immunity. 1998;9:677-686[CrossRef][Medline] [Order article via Infotrieve]. 10. Watt SM, Gschmeissner SE, Bates PA. PECAM-1: its expression and function as a cell adhesion molecule on hemopoietic and endothelial cells. Leuk Lymphoma. 1995;17:229-244[Medline] [Order article via Infotrieve]. 11. Shalaby F, Rossant J, Yamaguchi TP, et al. Failure of blood-island formation and vasculogenesis in Flk-1-deficient mice. Nature. 1995;376:62-66[CrossRef][Medline] [Order article via Infotrieve]. 12. Shalaby F, Ho J, Stanford WL, et al. A requirement for Flk1 in primitive and definitive hematopoiesis and vasculogenesis. Cell. 1997;89:981-990[CrossRef][Medline] [Order article via Infotrieve]. 13. Palis J, Robertson S, Kennedy M, Wall C, Keller G. Development of erythroid and myeloid progenitors in the yolk sac and embryo proper of the mouse. Development. 1999;126:5073-5084[Abstract].
14.
Palis J, Chan RJ, Koniski A, Patel R, Starr M, Yoder MC.
Spatial and temporal emergence of high proliferative potential hematopoietic precursors during murine embryogenesis.
Proc Natl Acad Sci U S A.
2001;98:4528-4533 15. Johnson GR, Moore MA. Role of stem cell migration in initiation of mouse foetal liver haemopoiesis. Nature. 1975;258:726-728[CrossRef][Medline] [Order article via Infotrieve]. 16. Houssaint E. Differentiation of the mouse hepatic primordium, II: extrinsic origin of the haemopoietic cell line. Cell Differ. 1981;10:243-252[CrossRef][Medline] [Order article via Infotrieve]. 17. Muller AM, Medvinsky A, Strouboulis J, Grosveld F, Dzierzak E. Development of hematopoietic stem cell activity in the mouse embryo. Immunity. 1994;1:291-301[CrossRef][Medline] [Order article via Infotrieve]. 18. Medvinsky A, Dzierzak E. Definitive hematopoiesis is autonomously initiated by the AGM region. Cell. 1996;86:897-906[CrossRef][Medline] [Order article via Infotrieve]. 19. Cumano A, Dieterlen-Lievre F, Godin I. Lymphoid potential, probed before circulation in mouse, is restricted to caudal intraembryonic splanchnopleura. Cell. 1996;86:907-916[CrossRef][Medline] [Order article via Infotrieve]. 20. Sanchez MJ, Holmes A, Miles C, Dzierzak E. Characterization of the first definitive hematopoietic stem cells in the AGM and liver of the mouse embryo. Immunity. 1996;5:513-525[CrossRef][Medline] [Order article via Infotrieve]. 21. Cumano A, Ferraz JC, Klaine M, Di Santo JP, Godin I. Intraembryonic, but not yolk sac hematopoietic precursors, isolated before circulation, provide long-term multilineage reconstitution. Immunity. 2001;15:477-485[CrossRef][Medline] [Order article via Infotrieve].
22.
Yoder MC, Hiatt K, Mukherjee P.
In vivo repopulating hematopoietic stem cells are present in the murine yolk sac at day 9.0 postcoitus.
Proc Natl Acad Sci U S A.
1997;94:6776-6780 23. Yoder MC, Hiatt K, Dutt P, Mukherjee P, Bodine DM, Orlic D. Characterization of definitive lymphohematopoietic stem cells in the day 9 murine yolk sac. Immunity. 1997;7:335-344[CrossRef][Medline] [Order article via Infotrieve].
24.
Matsuoka S, Tsuji K, Hisakawa H, et al.
Generation of definitive hematopoietic stem cells from murine early yolk sac and paraaortic splanchnopleures by aorta-gonad-mesonephros region-derived stromal cells.
Blood.
2001;98:6-12 25. Kyba M, Perlingeiro RC, Daley GQ. HoxB4 confers definitive lymphoid-myeloid engraftment potential on embryonic stem cell and yolk sac hematopoietic progenitors. Cell. 2002;109:29-37[CrossRef][Medline] [Order article via Infotrieve]. 26. Wiles MV, Keller G. Multiple hematopoietic lineages develop from embryonic stem (ES) cells in culture. Development. 1991;111:259-267[Abstract].
27.
Keller G, Kennedy M, Papayannopoulou T, Wiles MV.
Hematopoietic commitment during embryonic stem cell differentiation in culture.
Mol Cell Biol.
1993;13:473-486
28.
Nakano T, Kodama H, Honjo T.
Generation of lymphohematopoietic cells from embryonic stem cells in culture.
Science.
1994;265:1098-1101 29. Nakano T, Kodama H, Honjo T. In vitro development of primitive and definitive erythrocytes from different precursors. Science. 1996;272:722-724[Abstract].
30.
Vittet D, Prandini MH, Berthier R, et al.
Embryonic stem cells differentiate in vitro to endothelial cells through successive maturation steps.
Blood.
1996;88:3424-3431 31. Kennedy M, Firpo M, Choi K, et al. A common precursor for primitive erythropoiesis and definitive haematopoiesis. Nature. 1997;386:488-493[CrossRef][Medline] [Order article via Infotrieve]. 32. Choi K, Kennedy M, Kazarov A, Papadimitriou JC, Keller G. A common precursor for hematopoietic and endothelial cells. Development. 1998;125:725-732[Abstract]. 33. Perlingeiro RC, Kyba M, Daley GQ. Clonal analysis of differentiating embryonic stem cells reveals a hematopoietic progenitor with primitive erythroid and adult lymphoid-myeloid potential. Development. 2001;128:4597-4604[Medline] [Order article via Infotrieve]. 34. Porcher C, Swat W, Rockwell K, Fujiwara Y, Alt FW, Orkin SH. The T cell leukemia oncoprotein SCL/tal-1 is essential for development of all hematopoietic lineages. Cell. 1996;86:47-57[CrossRef][Medline] [Order article via Infotrieve]. 35. Robb L, Elwood NJ, Elefanty AG, et al. The scl gene product is required for the generation of all hematopoietic lineages in the adult mouse. EMBO J. 1996;15:4123-4129[Medline] [Order article via Infotrieve].
36.
Visvader JE, Fujiwara Y, Orkin SH.
Unsuspected role for the T-cell leukemia protein SCL/tal-1 in vascular development.
Genes Dev.
1998;12:473-479 37. Faloon P, Arentson E, Kazarov A, et al. Basic fibroblast growth factor positively regulates hematopoietic development. Development. 2000;127:1931-1941[Abstract].
38.
Phillips DR, Charo IF, Parise LV, Fitzgerald LA.
Platelet membrane glycoprotein IIb-IIIa complex.
Blood.
1988;71:831-843
39.
Wang Q, Stacy T, Binder M, Marin-Padilla M, Sharpe AH, Speck NA.
Disruption of the Cbfa2 gene causes necrosis and hemorrhaging in the central nervous system and blocks definitive hematopoiesis.
Proc Natl Acad Sci U S A.
1996;93:3444-3449
40.
Shivdasani RA, Orkin SH.
The transcriptional control of hematopoiesis.
Blood.
1996;87:4025-4039 41. Orkin SH, Porcher C, Fujiwara Y, Visvader J, Wang LC. Intersections between blood cell development and leukemia genes. Cancer Res. 1999;59:1784-1787. 42. Okuda T, van Deursen J, Hiebert SW, Grosveld F, Downing JR. AML1, the target of multiple chromosomal translocations in human leukemia, is essential for normal fetal liver hematopoiesis. Cell. 1996;4:321-330[CrossRef].
43.
Ogawa M, Kizumoto M, Nishikawa S, Fujimoto T, Kodama H, Nishikawa SI.
Expression of alpha4-integrin defines the earliest precursor of hematopoietic cell lineage diverged from endothelial cells.
Blood.
1999;93:1168-1177 44. Corbel C, Salaun J. AlphaIIb integrin expression during development of the murine hemopoietic system. Dev Biol. 2002;243:301-311[CrossRef][Medline] [Order article via Infotrieve]. 45. North TE, de Bruijn MF, Stacy T, et al. Runx1 expression marks long-term repopulating hematopoietic stem cells in the midgestation mouse embryo. Immunity. 2002;16:661-672[CrossRef][Medline] [Order article via Infotrieve].
46.
Ema H, Nakauchi H.
Expansion of hematopoietic stem cells in the developing liver of a mouse embryo.
Blood.
2000;95:2284-2288
47.
Debili N, Robin C, Schiavon V, et al.
Different expression of CD41 on human lymphoid and myeloid progenitors from adults and neonates.
Blood.
2001;97:2023-2030
48.
Ody C, Vaigot P, Quere P, Imhof BA, Corbel C.
Glycoprotein IIb-IIIa is expressed on avian multilineage hematopoietic progenitor cells.
Blood.
1999;93:2898-2906 49. Mitjavila-Garcia MT, Cailleret M, Godin I, et al. Expression of CD41 on hematopoietic progenitors derived from embryonic hematopoietic cells. Development. 2002;129:2003-2013[Medline] [Order article via Infotrieve].
50.
Morrison SJ, Hemmati HD, Wandycz AM, Weissman IL.
The purification and characterization of fetal liver hematopoietic stem cells.
Proc Natl Acad Sci U S A.
1995;92:10302-10306 51. Rebel VI, Miller CL, Thornbury GR, Dragowska WH, Eaves CJ, Lansdorp PM. A comparison of long-term repopulating hematopoietic stem cells in fetal liver and adult bone marrow from the mouse. Exp Hematol. 1996;24:638-648[Medline] [Order article via Infotrieve].
52.
Sato T, Laver JH, Ogawa M.
Reversible expression of CD34 by murine hematopoietic stem cells.
Blood.
1999;94:2548-2554 53. Emambokus NR, Frampton J. GPIIb is expressed on early multipotential haematopoietic progenitor cells and influences their number in the yolk sac, foetal liver and bone marrow [abstract]. Blood. 2001;98:711.
54.
Tronik-Le Roux D, Roullot V, Poujol C, Kortulewski T, Nurden P, Marguerie G.
Thrombasthenic mice generated by replacement of the integrin alpha(IIb) gene: demonstration that transcriptional activation of this megakaryocytic locus precedes lineage commitment.
Blood.
2000;96:1399-1408
55.
Emambokus N, Frampton J.
Analysis of GPIIb (
© 2003 by The American Society of Hematology.
| |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
 |
|
 |
|
  S. L. McKinney-Freeman, O. Naveiras, F. Yates, S. Loewer, M. Philitas, M. Curran, P. J. Park, and G. Q. Daley Surface antigen phenotypes of hematopoietic stem cells from embryos and murine embryonic stem cells Blood, July 9, 2009; 114(2): 268 - 278. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 T. Yamane, N. Hosen, H. Yamazaki, and I. L. Weissman Expression of AA4.1 marks lymphohematopoietic progenitors in early mouse development PNAS, June 2, 2009; 106(22): 8953 - 8958. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
C. Gekas, K. E. Rhodes, L. M. Gereige, H. Helgadottir, R. Ferrari, S. K. Kurdistani, E. Montecino-Rodriguez, R. Bassel-Duby, E. Olson, A. V. Krivtsov, et al. Mef2C is a lineage-restricted target of Scl/Tal1 and regulates megakaryopoiesis and B-cell homeostasis Blood, April 9, 2009; 113(15): 3461 - 3471. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 S. Irion, M.C. Nostro, S.J. Kattman, and G.M. Keller Directed Differentiation of Pluripotent Stem Cells: From Developmental Biology to Therapeutic Applications Cold Spring Harb Symp Quant Biol, March 27, 2009; (2009) sqb.2008.73.065v1. [Abstract] [PDF]
|
|
|
|
|
|
T. Yokota, K. Oritani, S. Butz, K. Kokame, P. W. Kincade, T. Miyata, D. Vestweber, and Y. Kanakura The endothelial antigen ESAM marks primitive hematopoietic progenitors throughout life in mice Blood, March 26, 2009; 113(13): 2914 - 2923. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M.-J. Yoon, B.-K. Koo, R. Song, H.-W. Jeong, J. Shin, Y.-W. Kim, Y.-Y. Kong, and P.-G. Suh Mind bomb-1 Is Essential for Intraembryonic Hematopoiesis in the Aortic Endothelium and the Subaortic Patches Mol. Cell. Biol., August 1, 2008; 28(15): 4794 - 4804. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 J. Y. Bertrand, A. D. Kim, S. Teng, and D. Traver CD41+ cmyb+ precursors colonize the zebrafish pronephros by a novel migration route to initiate adult hematopoiesis Development, May 15, 2008; 135(10): 1853 - 1862. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. L. McKinney-Freeman, C. Lengerke, I.-H. Jang, S. Schmitt, Y. Wang, M. Philitas, J. Shea, and G. Q. Daley Modulation of murine embryonic stem cell-derived CD41+c-kit+ hematopoietic progenitors by ectopic expression of Cdx genes Blood, May 15, 2008; 111(10): 4944 - 4953. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. Pearson, P. Sroczynska, G. Lacaud, and V. Kouskoff The stepwise specification of embryonic stem cells to hematopoietic fate is driven by sequential exposure to Bmp4, activin A, bFGF and VEGF Development, April 15, 2008; 135(8): 1525 - 1535. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 X. Zhao, V. Jankovic, A. Gural, G. Huang, A. Pardanani, S. Menendez, J. Zhang, R. Dunne, A. Xiao, H. Erdjument-Bromage, et al. Methylation of RUNX1 by PRMT1 abrogates SIN3A binding and potentiates its transcriptional activity Genes & Dev., March 1, 2008; 22(5): 640 - 653. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 A. Subramanian, S. Hegde, P. Porayette, M. Yon, P. Hankey, and R. F. Paulson Friend Virus Utilizes the BMP4-Dependent Stress Erythropoiesis Pathway To Induce Erythroleukemia J. Virol., January 1, 2008; 82(1): 382 - 393. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
H.-T. Huang and L.I. Zon Regulation of Stem Cells in the Zebra Fish Hematopoietic System Cold Spring Harb Symp Quant Biol, January 1, 2008; 73(0): 111 - 118. [Abstract] [PDF]
|
|
|
|
|
|
J. Y. Bertrand, A. D. Kim, E. P. Violette, D. L. Stachura, J. L. Cisson, and D. Traver Definitive hematopoiesis initiates through a committed erythromyeloid progenitor in the zebrafish embryo Development, December 1, 2007; 134(23): 4147 - 4156. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
P. Frontelo, D. Manwani, M. Galdass, H. Karsunky, F. Lohmann, P. G. Gallagher, and J. J. Bieker Novel role for EKLF in megakaryocyte lineage commitment Blood, December 1, 2007; 110(12): 3871 - 3880. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. Astorga and P. Carlsson Hedgehog induction of murine vasculogenesis is mediated by Foxf1 and Bmp4 Development, October 15, 2007; 134(20): 3753 - 3761. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
R. C. R. Perlingeiro Endoglin is required for hemangioblast and early hematopoietic development Development, August 15, 2007; 134(16): 3041 - 3048. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 H. K. Bone and M. J. Welham Phosphoinositide 3-kinase signalling regulates early development and developmental haemopoiesis J. Cell Sci., May 15, 2007; 120(10): 1752 - 1762. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. Kennedy, S. L. D'Souza, M. Lynch-Kattman, S. Schwantz, and G. Keller Development of the hemangioblast defines the onset of hematopoiesis in human ES cell differentiation cultures Blood, April 1, 2007; 109(7): 2679 - 2687. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 D. Rajesh, N. Chinnasamy, S. M. Mitalipov, D. P. Wolf, I. Slukvin, J. A. Thomson, and A. F. Shaaban Differential Requirements for Hematopoietic Commitment Between Human and Rhesus Embryonic Stem Cells Stem Cells, February 1, 2007; 25(2): 490 - 499. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 K. Kim, P. Lerou, A. Yabuuchi, C. Lengerke, K. Ng, J. West, A. Kirby, M. J. Daly, and G. Q. Daley Histocompatible Embryonic Stem Cells by Parthenogenesis Science, January 26, 2007; 315(5811): 482 - 486. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M. Bhatia Hematopoietic Development from Human Embryonic Stem Cells Hematology, January 1, 2007; 2007(1): 11 - 16. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 T. Umemoto, M. Yamato, Y. Shiratsuchi, M. Terasawa, J. Yang, K. Nishida, Y. Kobayashi, and T. Okano Expression of Integrin beta3 Is Correlated to the Properties of Quiescent Hemopoietic Stem Cells Possessing the Side Population Phenotype J. Immunol., December 1, 2006; 177(11): 7733 - 7739. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
H. K. A. Mikkola and S. H. Orkin The journey of developing hematopoietic stem cells. Development, October 1, 2006; 133(19): 3733 - 3744. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
L. Pang, H.-H. Xue, G. Szalai, X. Wang, Y. Wang, D. K. Watson, W. J. Leonard, G. A. Blobel, and M. Poncz Maturation stage-specific regulation of megakaryopoiesis by pointed-domain Ets proteins Blood, October 1, 2006; 108(7): 2198 - 2206. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
C. Rolny, I. Nilsson, P. Magnusson, A. Armulik, L. Jakobsson, P. Wentzel, P. Lindblom, J. Norlin, C. Betsholtz, R. Heuchel, et al. Platelet-derived growth factor receptor-beta promotes early endothelial cell differentiation Blood, September 15, 2006; 108(6): 1877 - 1886. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. A. Vodyanik, J. A. Thomson, and I. I. Slukvin Leukosialin (CD43) defines hematopoietic progenitors in human embryonic stem cell differentiation cultures Blood, September 15, 2006; 108(6): 2095 - 2105. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
H. Sakamoto, G. Dai, K. Tsujino, K. Hashimoto, X. Huang, T. Fujimoto, M. Mucenski, J. Frampton, and M. Ogawa Proper levels of c-Myb are discretely defined at distinct steps of hematopoietic cell development Blood, August 1, 2006; 108(3): 896 - 903. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
I. Kim, S. He, O. H. Yilmaz, M. J. Kiel, and S. J. Morrison Enhanced purification of fetal liver hematopoietic stem cells using SLAM family receptors Blood, July 15, 2006; 108(2): 737 - 744. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S.-i. Takayanagi, T. Hiroyama, S. Yamazaki, T. Nakajima, Y. Morita, J. Usui, K. Eto, T. Motohashi, K. Shiomi, K. Keino-Masu, et al. Genetic marking of hematopoietic stem and endothelial cells: identification of the Tmtsp gene encoding a novel cell surface protein with the thrombospondin-1 domain Blood, June 1, 2006; 107(11): 4317 - 4325. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
T. Yokota, J. Huang, M. Tavian, Y. Nagai, J. Hirose, J.-C. Zuniga-Pflucker, B. Peault, and P. W. Kincade Tracing the first waves of lymphopoiesis in mice Development, May 15, 2006; 133(10): 2041 - 2051. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
O. H. Yilmaz, M. J. Kiel, and S. J. Morrison SLAM family markers are conserved among hematopoietic stem cells from old and reconstituted mice and markedly increase their purity Blood, February 1, 2006; 107(3): 924 - 930. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
Z. Li, M. J. Chen, T. Stacy, and N. A. Speck Runx1 function in hematopoiesis is required in cells that express Tek Blood, January 1, 2006; 107(1): 106 - 110. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
Y. Wang, F. Yates, O. Naveiras, P. Ernst, and G. Q. Daley Embryonic stem cell-derived hematopoietic stem cells PNAS, December 27, 2005; 102(52): 19081 - 19086. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 A. Matsubara, A. Iwama, S. Yamazaki, C. Furuta, R. Hirasawa, Y. Morita, M. Osawa, T. Motohashi, K. Eto, H. Ema, et al. Endomucin, a CD34-like sialomucin, marks hematopoietic stem cells throughout development J. Exp. Med., December 5, 2005; 202(11): 1483 - 1492. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
H.-F. Lin, D. Traver, H. Zhu, K. Dooley, B. H. Paw, L. I. Zon, and R. I. Handin Analysis of thrombocyte development in CD41-GFP transgenic zebrafish Blood, December 1, 2005; 106(12): 3803 - 3810. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. Taoudi, A. M. Morrison, H. Inoue, R. Gribi, J. Ure, and A. Medvinsky Progressive divergence of definitive haematopoietic stem cells from the endothelial compartment does not depend on contact with the foetal liver Development, September 15, 2005; 132(18): 4179 - 4191. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
G. Keller Embryonic stem cell differentiation: emergence of a new era in biology and medicine Genes & Dev., May 15, 2005; 19(10): 1129 - 1155. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
T. M. Schlaeger, H. K. A. Mikkola, C. Gekas, H. B. Helgadottir, and S. H. Orkin Tie2Cre-mediated gene ablation defines the stem-cell leukemia gene (SCL/tal1)-dependent window during hematopoietic stem-cell development Blood, May 15, 2005; 105(10): 3871 - 3874. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 P. U. Magnusson, R. Ronca, P. Dell'Era, P. Carlstedt, L. Jakobsson, J. Partanen, A. Dimberg, and L. Claesson-Welsh Fibroblast Growth Factor Receptor-1 Expression Is Required for Hematopoietic but not Endothelial Cell Development Arterioscler. Thromb. Vasc. Biol., May 1, 2005; 25(5): 944 - 949. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. R. Gothert, S. E. Gustin, M. A. Hall, A. R. Green, B. Gottgens, D. J. Izon, and C. G. Begley In vivo fate-tracing studies using the Scl stem cell enhancer: embryonic hematopoietic stem cells significantly contribute to adult hematopoiesis Blood, April 1, 2005; 105(7): 2724 - 2732. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. A. Vodyanik, J. A. Bork, J. A. Thomson, and I. I. Slukvin Human embryonic stem cell-derived CD34+ cells: efficient production in the coculture with OP9 stromal cells and analysis of lymphohematopoietic potential Blood, January 15, 2005; 105(2): 617 - 626. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. Y. Bertrand, S. Giroux, R. Golub, M. Klaine, A. Jalil, L. Boucontet, I. Godin, and A. Cumano Characterization of purified intraembryonic hematopoietic stem cells as a tool to define their site of origin PNAS, January 4, 2005; 102(1): 134 - 139. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. J. Schuringa, K. Wu, G. Morrone, and M. A.S. Moore Enforced Activation of STAT5A Facilitates the Generation of Embryonic Stem-Derived Hematopoietic Stem Cells That Contribute to Hematopoiesis In Vivo Stem Cells, December 1, 2004; 22(7): 1191 - 1204. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
C. I. Baumann, A. S. Bailey, W. Li, M. J. Ferkowicz, M. C. Yoder, and W. H. Fleming PECAM-1 is expressed on hematopoietic stem cells throughout ontogeny and identifies a population of erythroid progenitors Blood, August 15, 2004; 104(4): 1010 - 1016. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
R. K. Burt, L. Verda, D.-A. Kim, Y. Oyama, K. Luo, and C. Link Embryonic Stem Cells As an Alternate Marrow Donor Source: Engraftment without Graft-Versus-Host Disease J. Exp. Med., April 5, 2004; 199(7): 895 - 904. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
W. Li, S. A. Johnson, W. C. Shelley, M. Ferkowicz, P. Morrison, Y. Li, and M. C. Yoder Primary endothelial cells isolated from the yolk sac and para-aortic splanchnopleura support the expansion of adult marrow stem cells in vitro Blood, December 15, 2003; 102(13): 4345 - 4353. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
D. Sugiyama, M. Ogawa, I. Hirose, T. Jaffredo, K.-i. Arai, and K. Tsuji Erythropoiesis from acetyl LDL incorporating endothelial cells at the preliver stage Blood, June 15, 2003; 101(12): 4733 - 4738. [Abstract] [Full Text] [PDF]
|
|
| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
 |
 |
 |
|||||||
 |
 |
 |
 |
 |
 |
 |
 |
||
| Copyright © 2003 by American Society of Hematology Online ISSN: 1528-0020 | |||||||||
Top
Abstract
Introduction
