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HEMATOPOIESIS
From Institut National de la Santé et de la
Recherche Médicale (INSERM) U 362, Institut Gustave Roussy,
Cedex, France.
The glycoprotein (Gp) IIb/IIIa integrin, also called CD41, is the
platelet receptor for fibrinogen and several other extracellular matrix
molecules. Recent evidence suggests that its expression is much wider
in the hematopoietic system than was previously thought. To investigate
the precise expression of the CD41 antigen during megakaryocyte (MK)
differentiation, CD34+ cells from cord blood and mobilized
blood cells from adults were grown for 6 days in the presence of stem
cell factor and thrombopoietin. Two different pathways of
differentiation were observed: one in the adult and one in the neonate
cells. In the neonate samples, early MK differentiation proceeded from
CD34+CD41 Hematopoietic stem cells are a heterogeneous
population of cells defined by both their multilineage potential and
their hematopoietic reconstitution capacities after
transplantation.1,2 In mice, biological assays have been
developed to evaluate the repopulating capacity of a cell population
and consequently its content in stem cells.3 Several in
vitro assays have been designed in parallel to better characterize the
hierarchy of hematopoietic primitive cells, which include true stem
cells and early multipotential progenitors. One of these is the
long-term culture initiating cell (LTC-IC) assay, which defines a
population of primitive progenitors close to true hematopoietic stem
cells.4-7 More recently, xenograft assays have been
developed to test the engrafting capacities of human cells although the
precise relationship between nonobese diabetic/severe combined
immune-deficient (NOD-SCID) repopulating cells and long-term
reconstituting human stem cells is still unknown.8-12
Definition of the hematopoietic hierarchy has been facilitated by the
identification of differentiation antigens.13-16 It has been generally considered that primitive hematopoietic cells lack lineage markers, assuming that they appear after commitment or during
late stages of differentiation. The glycoprotein (Gp) IIb integrin
( In this study, we have shown that CD41 antigen is expressed on a
fraction of LTC-ICs and NOD-SCID reconstituting cells as well as on
cells with a B-, T-, and natural killer (NK)-lymphoid potential
derived from cord blood cells. In adults, CD41 expression is more
restricted and is essentially present on erythroid and MK progenitors,
although the CD41+ cell population may include some
LTC-ICs. CD42 (GpIbIX), which is considered to be a later marker of the
megakaryocytic differentiation than CD41, is also detected on non-MK
progenitors from cord blood.
Antibodies
Unconjugated Tab mAb (anti-CD41) was a generous gift from R. Mac
Ever (Oklahoma City, OK). Alkaline-phosphatase-coupled
polyclonal goat antibody against mouse immunoglobulin (Caltag) was
purchased from Tebu (Le Perray-en-Yvelines, France).
Purification of CD34+ cells
In vitro liquid cultures of MKs from CD34+ cells CD34+ cells were grown for 6 days in serum-free Iscove's modified Dulbecco's medium (Gibco, Paisley, Scotland), prepared as previously reported.31 The medium was supplemented with a combination of pegylated recombinant human MK growth and development factor (PEG-rHuMGDF 10 ng/mL; a generous gift from Kirin, Tokyo, Japan) and 50 ng/mL recombinant human stem cell factor (SCF) (a generous gift from Amgen, Thousand Oaks, CA).Cell sorting of different CD41 subsets MKs at different stages of differentiation were obtained after 6 days of culture. Cells were incubated with a mixture of a FITC anti-CD42a, R-PE anti-CD41a, and R-PE-Cy5 anti-CD34 mAbs for 30 minutes at 4°C in their culture medium. Cells were washed in culture medium and sorted according to their immunophenotype into 6 different populations as follows: CD34+CD41a CD42a,
CD34+CD41a+CD42a,
CD34+CD41a+CD42a+,
CD34CD41aCD42a,
CD34CD41a+CD42a, and
CD34CD41a+CD42a+, by means
of a FACS Vantage cytometer (Becton Dickinson) equipped with an argon
laser (Coherent Radiation, Palo Alto, CA) and a 100-µm nozzle. Each
cell fraction was resorted in order to get a purity of greater
than 97%.
Quantification of clonogenic progenitors in semisolid cultures Serum-free fibrin clot assays.
Cultures were performed in serum-free fibrin clot assays in the
presence of cytokines. Ingredients for serum-free cultures were similar
to those of liquid cultures in which were added bovine plasma
fibrinogen (1 mg/mL) (Sigma, St Louis, MO), 0.01 M Methylcellulose assays. Erythroid (BFU-E) and granulocytic (CFU-granulo-monocytic [GM]) progenitors were quantified by means of previously described methylcellulose assays.32 Cultures were stimulated by addition of recombinant human (rHu) growth factors: PEG-rHuMGDF (10 ng/mL), SCF (50 ng/mL), G-CSF (20 ng/mL), IL-6 (100 U/mL), IL-3 (100 U/mL), and human Epo (1 U/mL). Hematopoietic progenitors were scored on day 12 by means of an inverted microscope. Detection of LTC-ICs in the sorted fractions The presence of LTC-ICs was assessed by culturing the sorted fractions on the murine MS-5 stromal cells as previously described.33 Cultures were initiated at limiting dilutions by plating 10 to 100 cells per well (96-well plates). Wells were maintained at 33°C, 5% CO2, and fed weekly by half medium change. The content in clonogenic progenitors of each well was assessed after 6 weeks in culture by plating nonadherent and adherent cells (recovered by trypsinization) in methylcellulose assay (see above) supplemented with IL-3, Epo, SCF, and G-CSF. We tested 20 wells per cell concentration of different cell fractions. A positive well was defined as a well that contained at least one clonogenic progenitor cell after 6 weeks in culture.Simultaneous assessment of erythroid, MK, and granulo-monocytic differentiation Cells from the different cell fractions were sorted individually by means of the automatic cloning design of the flow cytometer into 96-well tissue-culture plates. The medium was serum-free and contained a combination of 6 cytokines (SCF, IL-3, Epo, G-CSF, IL-6, and PEG-rHuMGDF).34 Plates were examined at days 11 to 13 and days 18 to 20 after incubation at 37°C in an air atmosphere supplemented with 5% CO2. Individual clones were analyzed by flow cytometry.Simultaneous assessment of B, NK, and granulo-monocytic differentiation The different cell fractions were incubated in 24- and 96-well plates precoated with confluent murine MS-5 cells in RPMI supplemented with 10% human serum, 5% fetal calf serum (FCS), and a combination of 7 cytokines (10 ng/mL IL-3; 50 ng/mL SCF; 50 ng/mL fetal liver tyrosine kinase-3 ligand [FLT3-L], a generous gift from Immunex; 10 ng/mL PEG-rHuMGDF; 5 ng/mL IL-2; 10 ng/mL IL-15; and 20 ng/mL IL-7) (the last 3 cytokines were from Diaclone). Wells with significant cell proliferation were collected after 4 to 6 weeks, and cell phenotype was determined by flow cytometry.35Assessment of T-cell potential in fetal thymus organ culture (FTOC) Isolation of murine embryonic NOD-SCID thymic lobes, incubation with human cells by means of the hanging drop procedure, and organotypic cultures were performed following standard procedures initially described to analyze mouse T-lymphoid differentiation and adapted to the identification of human T-cell potential.36 The standard technique has been slightly modified by adding a combination of cytokines (5 ng/mL IL-2; 20 ng/mL IL-7; and 50 ng/mL SCF) during the hanging drop procedure.35 Cells recovered from the thymic lobes after about 30 days were studied by immunofluorescence and analyzed by flow cytometry.35Assessment of the ability of hematopoietic cells to engraft NOD-SCID NOD/LtSz-scid /scid mice were irradiated with a single dose of 300 cGy from an x-ray source (Philips, Eindhoven, The Netherlands) before transplantation. Mice were anesthesized shortly thereafter with ether, and cultured cells of different phenotypes were injected in the retro-orbital vein. Long bones from the recipient mice were analyzed after 5 to 8 weeks by flow cytometry. A mouse was considered positive if at least 0.1% human cells (CD45+) were detected in comparison with the isotype control.Immunolabeling for flow cytometric analysis Individual clones were simultaneously labeled with a PerCP anti-CD61, R-PE anti-GPA, and FITC anti-CD15 mAbs to estimate erythroid, MK, and granulo-monocytic differentiation; Cy5/R-PE anti-CD56, R-PE anti-CD19, and FITC anti-CD15 mAbs for evaluation of B, NK, and granulo-monocytic differentiation; and R-PE anti-human CD4 and FITC anti-human CD8 in FTOC assays. These 2 antibodies recognized exclusively human cells.The presence of human cells in NOD/SCID mouse bone marrow was studied after labeling with a FITC-anti-human CD45 mAb. To determine the precise phenotype of human cells, double or triple staining was performed by means of the FITC anti-CD45 mAb, FITC anti-CD15, an R-PE-Cy5 anti-CD34 mAb, and an R-PE anti-CD19, anti-CD11b, or anti-CD41 mAb. The phenotype was determined in a gate that was the intersection of a morphological (scatter properties) and a CD45+ gate. Cell samples were analyzed on a FACSort (Becton Dickinson). Cells were analyzed with the Cellquest software package (Becton Dickinson).
Immunophenotypic characterization of CD41+ cells deriving from adult and cord blood CD34+ cells CD34+ cells from cord blood or leukapheresis were cultured in the presence of SCF plus PEG-rHuMGDF. As previously demonstrated,37,38 a high percentage of CD41+ cells were present after 6 days of culture with a wide diversity in its expression. Triple staining with anti-CD34, anti-CD41, and anti-CD42 antibodies was performed as illustrated in Figure 1A-C. A direct linear relationship between expression of CD41 and CD42 was observed (Figure 1A). Thus, only cells expressing a low level of CD41 (CD41+) were devoid of CD42 (see gate R2). In contrast, intermediate or high expression of CD41 (CD41++) correlated with the presence of CD42 (see gate R3). These results were obtained with both neonate and adult cells. The CD41+CD42 cells include both
CD34+ and CD34 cells. CD34+ cells
co-expressing both CD41 and CD42
(CD34+CD41++CD42+) were also
detected. Striking differences were observed between adult and neonate
cultures because at day 6 the great majority of CD41+ cells
were CD34+ in the adult culture (Figure 1B, gates R2 + R3) and CD34 in the neonate culture (Figure 1A, gates
R2 + R3). This suggests that differentiation may proceed along
different pathways in the adult and the neonate.
To demonstrate this hypothesis, kinetics were performed, and cells were
labeled every day starting on the day of purification to day 6 (Figure
1B). On freshly purified cells, the precise percentage of
CD34+CD41+ cells was sometimes difficult to
determine owing to platelet fragment binding, which could not be
totally eliminated either by neuraminidase or elastase treatment. From
day 2 to day 3, the percentage of CD41+ cells derived from
cord blood was low (up to 10%) and was present on CD34+
cells with an intermediate level of antigen. At day 4, the
CD41+ cells switched to a CD34 Thus, different pathways of early megakaryocytic differentiation
occurred in the adult and the neonate cultures, as illustrated schematically in Figure 1C, and leads to a common mature phenotype (CD34 The percentage of cells expressing the different antigenic
combinations at day 6 varied from one experiment to another, but differences between adult and neonate cultures were constantly observed, as shown in Figure 1A (n = 15). In cord blood cultures, the
percentage of CD34+CD41+CD42 To precisely determine the properties of those cells, the different cell fractions were sorted at day 6 of culture and studied by biological assays. Expression of the CD41 antigen on hematopoietic clonogenic progenitors Cells were sorted according to the expression of CD34, CD41, and CD42 into 6 cell fractions (CD34+CD41aCD42a,
CD34+CD41a+CD42a,
CD34CD41aCD42a,
CD34CD41a+CD42a,
CD34+CD41a++CD42a+, and
CD34CD41a++CD42a++) at day 6 of
culture and grown in methylcellulose in the presence of 5 cytokines to
reveal their erythroid and myeloid potential and in plasma clot to
reveal their erythro-MK potential. Results of 10 experiments performed
from both mobilized adult blood and cord blood cells are summarized in
Figure 2. Cloning efficiency of CFU-GM,
BFU-E, CFU-MK, and BFU-E/MK was quite similar in the CD34+CD41CD42 cell fractions
from adult and cord blood samples (219.4/103,
112.7/103, 45.4/103, and 26.3/103
for BFU-E, CFU-GM, CFU-MK and BFU-E/MK for leukapheresis vs
184.4/103, 107/103,
34.7/103, and 15.6/103 for cord
blood). Thus, cloning efficiency in each fraction was referred to the cloning efficiency of this cell fraction. In the adult
blood, non-MK progenitors were essentially found in the CD34+CD41CD42 cell population.
However, some BFU-E and CFU-GM (less than 20% of the
CD34+CD41CD42 cell cloning
efficiency) were found in the 2 cell fractions expressing CD41 but not
CD42, irrespective of CD34 expression. Surprisingly, progenitors were
also present in the
CD34CD41CD42 cell fraction,
especially CFU-GM (50% of the
CD34+CD41CD42 cell cloning
efficiency). In cord blood, non-MK progenitors have a higher cloning
efficiency in all the CD41+ cell fractions except the
CD34CD41+CD42+ cell fractions
than in the adult blood. Furthermore, non-MK progenitors, essentially
erythroid progenitors, were present in cell fractions expressing CD42.
All these differences were statistically significant, and cloning
efficiency of the CD34+CD41+CD42
cell population for BFU-E and CFU-GM was about 60% that found in the
CD34+CD41CD42 cell fraction. In
contrast to the adult blood cells, BFU-E were enriched in the cultured
CD34CD41CD42 cell fraction.
Differences were also observed for MK progenitors. Indeed, MK
progenitors and the erythro/MK progenitor (BFU-E/MK) were significantly
enriched in the
CD34+CD41+CD42+ and
CD34CD41+CD42 cell fractions in
comparison with the adult blood cells. In both the adult and neonate
blood cells, MK colonies derived from the fractions expressing CD42
were composed of very few cells.
To determine the myeloid potential of these CD41+ cells in more detail, limiting dilution experiments were performed with cord blood or adult cells. Expression of CD41 antigen on pluripotent myeloid progenitors Cells from the CD34+CD41 and
CD34+CD41+CD42 cell fractions
were deposited at one cell per well by means of the automated cell deposition unit of the flow cytometer and cultured from 10 to 13 days.
Each well, which comprises more than 200 cells, was immunophenotyped by
triple staining. This procedure enabled us to define precisely the MK
(CD61high cells with high FCS and SSC), erythroid (GPA),
and granulo-monocytic (CD15) potential of each clone. Results of cord
blood cultures obtained from 4 separate experiments and derived from
112 and 95 wells are illustrated in Figure
3. Cloning efficiency was slightly lower
in the CD34+CD41+CD42 cell
population than in the CD34+CD41 (35% vs
28%). All potentialities could be found in the
CD34+CD41+ cells, including progenitors with a
erythro/granulo-monocytic/MK potential. It must be emphasized that in
these experiments the frequency of MK progenitors was greatly
underestimated owing to the number of cells required for the flow
cytometric analysis (more than 200 cells). In contrast, in the adult,
almost all clones derived from
CD34+CD41+CD42 and containing
more than 200 cells were purely erythroid or mixed erythro/MK. The
cloning efficiency was low (about 5%) in the
CD34+CD41+CD42 cell population as
compared with the CD34+CD41 (32%) (data
not shown).
Expression of CD41 antigen on primitive hematopoietic cells (LTC-ICs and NOD-SCID repopulating cells) Consequently, we investigated whether the CD41 antigen was expressed on primitive hematopoietic cells revealed by the LTC-IC assay. LTC-ICs were detected in the CD34+CD41CD42 and
CD34+CD41+CD42 cell fractions but
not in the CD34CD41+CD42
cell subset in samples from cultured cord blood and adult mobilized CD34+ cells. The LTC-IC frequency was reduced about 4- and
2-fold in the CD34+CD41+CD42 cell
fraction in comparison with the
CD34+CD41 cell fraction from the adult and
neonate samples, respectively (1/38 vs 1/138; 1/30 vs 1/65) (Figure
4A). In absolute number, when the
frequency of each cell fraction was taken into account, the LTC-ICs
contained in the adult or neonate
CD34+CD41+CD42 cell subset in
comparison with those contained in the
CD34+CD41 cell fraction was in the same order
of magnitude (8.9% in the neonate and 8.5% in the adult
cells).
In order to determine if the cells bearing the CD41 antigen may be
transplantable in vivo, 3 cord blood cell fractions
(CD34+CD41 Phenotypic analysis of the human hematopoietic cells reconstituted with these 2 cell fractions was not markedly different, and CD19+ cells were the predominant cell population in the mice (Figure 4B). This clearly suggests that CD34+CD41+ cells also have a B-lymphoid potential. NOD-SCID repopulating experiments were performed with mobilized blood
CD34+ in 2 experiments. The level of reconstitution was low
at the threshold of detection (0.8%) with both cell fractions
(CD34+CD41 B-cell and NK-cell differentiation potential of CD41+ cells To more precisely ascertain that CD41 antigen may be expressed on lymphoid progenitors, cord blood CD34+CD41
and CD34+CD41+CD42 cells were
grown on the murine stromal cell line MS-5 in the presence of a
combination of 7 cytokines (SCF, IL-3, FLT3-L, PEG-2HumGDF, IL-15,
IL-7, and IL-2) for 4 to 6 weeks. Experiments were performed at one
cell per well, comparing the CD34+CD41 and
the entire CD34+CD41+ cell fractions. Frequency
of proliferating cells (more than 200 cells) was 3-fold less in the
CD34+CD41+ cell fraction than in the
CD34+CD41 cell fraction (58 of 1080 vs 180 of
1068) at 4 or 6 weeks. These proliferating clones were
immunophenotyped. All types of potentialities except pure B cells were
found in the CD34+CD41+ cell fractions,
including cells with the 3 potentialities (NK/M/B) (Figure
5). In the adult cells, similar
experiments were performed. However, the cloning efficiency of
CD34+CD41+CD42 cells in
lympho-myeloid conditions was extremely low (about 0.3% on 2000 clones
studied), and wells contained only myeloid cells (CD15high).
T-lymphoid potential of CD41+ cells These experiments demonstrated that the CD41 antigen is present on cord blood cells with a lymphoid potential (B and NK). However, it remained to be determined if the CD34+CD41+ also had a T-cell potential. NOD-SCID embryonic thymus can be used to reveal the T-cell potential of human CD34+ cells.35,40 Using this strategy, we investigated the T-cell potential of the CD34+CD41,
CD34+CD41+CD42, and
CD34CD41+CD42 cord blood cells.
First, 25 000 cells were used in the hanging drop procedure; cells
were recovered after 4 weeks of culture and characterized with
antibodies against human CD4 and CD8. In these conditions,
CD34+CD41CD42 and
CD34+CD41+CD42 cells generated T
cells (CD4+ and double-positive
CD4+/CD8+ cells) in all thymic lobes whereas
the CD34CD41+CD42 cell fraction
did not have this potential (Figure 6A).
Limiting dilution experiments were then performed with the 2 first
fractions and revealed that the frequency of cells with a T-cell
potential was quite similar in the
CD34+CD41CD42 and
CD34+CD41+CD42 cells (Figure 6B).
These experiments were not performed with adult cells.
Hematopoietic stem cells are a heterogeneous population of cells
that are able to reconstitute hematopoiesis. Numerous differentiation membrane antigens have been defined to better identify this cell population. Among them, CD34 is the most widely used antigen to characterize hematopoietic stem cells and progenitors. However, a
fraction of stem cells that are considered extremely primitive are
CD34 Expression of CD41 in the hematopoietic system is controversial.21,22,24,27-30,45-47 It is well accepted that CD41 is detected during MK differentiation at a stage of a late MK progenitor.21,45,47 The presence of CD41 on multipotent progenitors and some non-MK-committed progenitors is still a matter of debate.22-24,48 This controversy might be due to the level of the CD41 antigen expressed on different cell types. A very high expression is present during late MK differentiation whereas a low amount of the protein might be present on other lineages as suggested by the work of Tropel et al.28 To obtain a high number of CD41+ cells, we have cultured purified CD34+ cells in the presence of a combination of SCF and PEG-rHuMGDF, a condition that induced both MK differentiation and expansion of hematopoietic primitive cells.31,49 Previously, Basch et al48 have shown that TPO could induce CD41 expression on adult CD34+ cells after a 24-hour incubation. In some cell lines, it has been also shown that TPO increases synthesis of CD41.50 However, in normal cells, induction of the CD41 antigen on CD34+ cells is not a restricted effect of TPO because IL-3 or a combination of several cytokines had the same effects.37,48 Furthermore, there is evidence that cells with induced CD41 antigen have the same properties as freshly isolated cells.48 There remain several advantages to culturing CD34+ cells for studying CD41 expression: (1) the development of a large number of CD41+ cells; (2) the possibility of analyzing rare CD41+ phenotypes; and (3) minimizing contamination by platelet fragments stuck to CD34+ cells during cell purification. When cultured cells were stained with antibodies against CD42 and CD41,
a linear relationship between expression of these 2 antigens was
observed. Only cells expressing low levels of CD41 were
CD42 These immunophenotypic differences correlate with different ontogenic
biological properties of the CD41+ cells. In cord blood,
the CD34+CD41+CD42 Surprisingly, these ontogenic changes may also involve the CD42 antigen. In neonate culture, expression of CD42 on CD34+CD41+ cells did not indicate MK commitment since erythroid and myeloid progenitors were detected in this cell fraction, as previously suggested.54 The present study further emphasizes the ontogenic changes occurring in
the hematopoietic stem cell compartment and during MK differentiation.
We observed 2 different pathways of MK differentiation with respect to
surface markers in the adult and neonate samples (Figure 1C). This
difference involves mainly CD34 expression. MK commitment occurs in a
CD34
The authors thank P. Ardouin and A. Rouchez for breeding and care of the NOD-SCID mice, Aline Massé for technical assistance in handling and phenotyping the NOD-SCID mice and Drs Catherine Bocaccio and Françine Norol for providing leukapheresis samples.
Submitted May 22, 2000; accepted November 29, 2000.
Supported by grants from the Institut National de la Santé et de la Recherche Médicale, Association pour la Recherche sur le cancer (ND grant 9728), and the Institut Gustave Roussy.
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: Najet Debili, INSERM U 362, Institut Gustave Roussy, Villejuif 94805, Cedex, France; e-mail: verpre{at}igr.fr.
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© 2001 by The American Society of Hematology.
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  R. Chaligne, C. James, C. Tonetti, R. Besancenot, J. P. Le Couedic, F. Fava, F. Mazurier, I. Godin, K. Maloum, F. Larbret, et al. Evidence for MPL W515L/K mutations in hematopoietic stem cells in primitive myelofibrosis Blood, November 15, 2007; 110(10): 3735 - 3743. [Abstract] [Full Text] [PDF]
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F. Delhommeau, S. Dupont, C. Tonetti, A. Masse, I. Godin, J.-P. L. Couedic, N. Debili, P. Saulnier, N. Casadevall, W. Vainchenker, et al. Evidence that the JAK2 G1849T (V617F) mutation occurs in a lymphomyeloid progenitor in polycythemia vera and idiopathic myelofibrosis Blood, January 1, 2007; 109(1): 71 - 77. [Abstract] [Full Text] [PDF]
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M. C. Martinez, F. Larbret, F. Zobairi, J. Coulombe, N. Debili, W. Vainchenker, M. Ruat, and J.-M. Freyssinet Transfer of differentiation signal by membrane microvesicles harboring hedgehog morphogens Blood, November 1, 2006; 108(9): 3012 - 3020. [Abstract] [Full Text] [PDF]
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B. Jacquelin, T. Kortulewski, P. Vaigot, A. Pawlik, G. Gruel, O. Alibert, P. Soularue, C. Joubert, X. Gidrol, and D. T.-L. Roux Novel pathway for megakaryocyte production after in vivo conditional eradication of integrin {alpha}IIb-expressing cells Blood, September 15, 2005; 106(6): 1965 - 1974. [Abstract] [Full Text] [PDF]
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J.-M. Paulus, N. Debili, F. Larbret, J. Levin, and W. Vainchenker Thrombopoietin responsiveness reflects the number of doublings undergone by megakaryocyte progenitors Blood, October 15, 2004; 104(8): 2291 - 2298. [Abstract] [Full Text] [PDF]
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M. T. Mitjavila-Garcia, M. Cailleret, I. Godin, M. M. Nogueira, K. Cohen-Solal, V. Schiavon, Y. Lecluse, F. Le Pesteur, A. H. Lagrue, and W. Vainchenker Expression of CD41 on hematopoietic progenitors derived from embryonic hematopoietic cells Development, March 6, 2003; 129(8): 2003 - 2013. [Abstract] [Full Text] [PDF]
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H. K. A. Mikkola, Y. Fujiwara, T. M. Schlaeger, D. Traver, and S. H. Orkin Expression of CD41 marks the initiation of definitive hematopoiesis in the mouse embryo Blood, January 15, 2003; 101(2): 508 - 516. [Abstract] [Full Text] [PDF]
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R. L. Darley, L. Pearn, N. Omidvar, M. Sweeney, J. Fisher, S. Phillips, T. Hoy, and A. K. Burnett Protein kinase C mediates mutant N-Ras-induced developmental abnormalities in normal human erythroid cells Blood, December 1, 2002; 100(12): 4185 - 4192. [Abstract] [Full Text] [PDF]
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M. G. Manz, T. Miyamoto, K. Akashi, and I. L. Weissman Prospective isolation of human clonogenic common myeloid progenitors PNAS, September 3, 2002; 99(18): 11872 - 11877. [Abstract] [Full Text] [PDF]
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Top
Abstract
Introduction
IIb, CD41b) associates with GpIIIa (
3, CD61) to form a complex
(GpIIb/IIIa, CD41a) that functions as the fibrinogen receptor in
platelets.
amino caproic acid, and horse thrombin (6 × 10
