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CLINICAL HEMATOLOGY
ENRICO E. TUY, MD, FPCP, FPSHBT
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COURSE OBJECTIVES
• To acquire knowledge in the basic principles
underlying normal and abnormal hematologic
phenomena.
• To understand blood disorders in the context of thestructural and functional features of the
hematopoietic-lymphoid system.
• To apply principles of basic sciences (biochemistry,
immunology, molecular biology and genetics) in
understanding blood disorders
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References
• Hematology in Clinical Practice; Hillman RS et al(Paperback)
McGraw-Hill Professional; 4th edition (June 1, 2005)
• Essential Haematology; Hoffbrand V et al, Wiley-Blackwell;
6th
edition (2011)• Harrison’s Textbook of Internal Medicine; Fauci et al (eds);
McGraw Hill, 18th edition, 2012
• Haematology An Illustrated Colour Text; Howard MR,
Hamilton PJ, 3rd
Ed (2008) Churchchill Livingstone Elsevier
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INTRODUCTION
THE BLOOD
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Composition of the Blood
• Cellular element
– Red blood cells
– White blood cells
•Granulocytes
– Neutrophils
– Eosinophils
– Basophils
• Monocytes
• Lymphocytes
– Platelets
• Fluid elements
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RBC’s
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NEUTROPHILS
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BASOPHILS and EOSINOPHILS
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MONOCYTES
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MONOCYTES
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LYMPHOCYTES
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PLATELETS
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Composition of the Blood
• Fluid elements
– Water
– Electrolytes
– Proteins• Albumin
• Immunoglobulins and
complements
• Transport proteins
– Transferrin
– Transcobalamin
– Haptoglobin Serum electrophoresis
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HEMATOPOIESIS
Origin and Development
of Blood Cells
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Hematopoiesis
• Refers to the process of formation,
development, and differentiation of the
formed elements.
• The process originates from the
hematopoietic stem cells (HSCs)
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Hematopoietic Stem Cells (HSCs)
• Essential Attributes – Self-renewal
– Mutilineage differentiation
– Reconstitution of hematopoietic system of a lethallyirradiated host
– HSCs are rare, occurring at a frequency of 1 stem cellper 10,000 to 100,000 bone marrow (BM) cells.
– Under steady state, HSCs are quiescent (noncycling)
and only a fraction enter into the cell cycle. – Deregulation of hematopoiesis may lead to
malignancies or bone marrow failure syndromes
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Stem and Progenitor CellsCharacteristic Stem Cell Progenitor Cell
Proliferative potential Tremendous More limited
Renewal On a population basis Probably none
Potential for differentiation All lineages Restricted
Differentiated characteristi Minimal-lineage negative Progressively increases
Functional characteristics Rapid directed motility andexpression of proteopodia;
homing
Less defined
Cycle status Dormant Cycling
Cytokine responsiveness Large number of cytokines
needed
Restricted
Cell of origin Unknown Stem cell
Rhodamine and Hoechst
dyes stain
Stains dimly Bright
Adheres to marrow stroma Yes No or limited
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HEMATOPOIETIC STEM CELLS
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Figure 20-03
Copyright © 2005 Elsevier Inc. (USA) All rights reserved.
Site of Hematopoiesis
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Marrow Structure
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Bone Marrow Microenvironment
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Marrow Microenvironment
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Marrow Microenvironment
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Role of Marrow Microenvironment in
Hematopoiesis
• Provide a source of growth factors
• Important in the “trafficking” of immature
HSCs into and out of the marrow and the
blood. (Homing and Mobilization)
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Figure 17-01
Copyright © 2005 Elsevier Inc. (USA) All rights reserved.
The Hematopoietic Hierarchy
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Characterization of HSCs
• Surface markers
• Dye exclusion
•
In vitro culture• Enumeration of HSCs by transplantation
models
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Sources of HSCs
• Bone marrow
• The peripheral circulation after mobilization
by chemotherapy and cytokines (mPB),
• Umbilical cord blood
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Hematopoiesis
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Operational Components of
Hematopoiesis
• Anatomical structure of the marrow
•
The stroma or marrow microenvironment
• Hematopoietic stem cells
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HEMATOPOIETIC STEM AND PROGENITOR CELLS
Characteristic Stem Cell Progenitor Cell
Proliferative potential Large More limited
Renewal Yes Probably none
Potential for differentiation All lymphohematopoietic
lineages
Restricted
Differentiated
characteristics
Minimal-lineage negative Progressively increases to
lineage specific cell
Cycle status Dormant Cycling
Cytokine responsiveness Large number of cytokines
needed for expression of
phenotype
Restricted
Cell of origin Unknown Stem cell
Adheres to marrow stroma Yes Limited to none
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The Humoral Regulation of
Hematopoiesis
Cytokines
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Characteristics of Cytokines
• Are glycoproteins
• Act on cell surface receptors
• Initiate complex second messenger and post-translational regulation
• May act on stem cells, progenitor cells, anddifferentiated cells of the same lineage
• May act on multiple different lineages
• Stimulate or inhibit proliferation, apoptosis,
differentiation, or function• Usually act on neoplastic counterpart of normal target
cell
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Copyright ©2008 American Society of Hematology. Copyright restrictions may apply.
Metcalf, D. Blood 2008;111:485-491
No Caption FoundVarying Tissue Origin of Cytokines
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Copyright ©2008 American Society of Hematology. Copyright restrictions may apply.
Metcalf, D. Blood 2008;111:485-491
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Copyright ©2008 American Society of Hematology. Copyright restrictions may apply.
Metcalf, D. Blood 2008;111:485-491
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Copyright ©2008 American Society of Hematology. Copyright restrictions may apply.
Metcalf, D. Blood 2008;111:485-491
No Caption Found
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Cytokines with Hematopoietic
Activities
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Colony-stimulating factors
• Granulocyte colony-stimulating factor (G-CSF)
• Granulocyte-macrophage colony-stimulatingfactor (GM-CSF)
• Macrophage colony-stimulating factor (M-CSF)
• Interleukin-3 (IL-3)
• Erythropoietin (EPO)
• Thrombopoieitin (TPO)
• Interleukin-5 (IL-5)
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Figure 19-01
Copyright © 2005 Elsevier Inc. (USA) All rights reserved.
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HEMATOPOIETIC PROGENITORS ASSAYED IN VITRO
Cell Type Lineage Primary Cytokines
CFU-GEMM Neutrophil, erythroid, macrophage, T
cell, megakaryoctes
IL-3,GM-CSF, EPO
CFU-GM Neutrophil, monocytes-macrophages GM-CSF
CFU-M Monocyte-macrophage M-CSF (CSF-1)
CFU-G Neutrophil G-CSF
CFU-EOS Eosinophil IL-5, IL-3, GM-CSF
BFU-E Erythroid IL-3, KIT Ligand, GM-CSF,
IL-4, EPO
CFU-E Erythroid EPO
BFU-meg Megakaryocytes IL-3, GM-CSF,c-mpl ligand
CFU-meg Megakaryoctes IL-3, GM-CSF, c-mpl ligand,
IL-11, IL-6, LIF, G-CSF, KIT
ligand, IL-4
CFU-baso Mast cell, basophil IL-3, KIT ligand, GM-CSF
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Table 19-01
Copyright © 2005 Elsevier Inc. (USA) All rights reserved.
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Table 19-02
Copyright © 2005 Elsevier Inc. (USA) All rights reserved.
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The Bone Marrow
The Hematopoietic
Microenvironment
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Cellular Components of the Marrow
Microenvironment
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• Metcalf , D Hematopoietic cytokines Blood, 15 January 2008, Vol. 111, No. 2, pp. 485-491.
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“THROUGHOUT THE CENTURIES
THERE WERE MEN (WOMEN)
WHO TOOK FIRST STEPS
DOWN NEW ROADS
ARMED WITH NOTHINGBUT THEIR OWN VISION”
AYN RAND
THE FOUNTAINHEAD
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HEMATOPOIESIS II
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Erythropoiesis (Progenitor Compartment)
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Erythropoiesis (Precursor Compartment
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Granulocytopoiesis
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Granulocyte Maturation
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Thrombocytopoiesis
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CLINICAL HEMATOLOGY
ENRICO E. TUY, MD, FPCPDiplomate Philippine Society of Hematology & Blood
Transfusion
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COURSE OBJECTIVES
• To acquire knowledge in the basic principles
underlying normal and abnormal hematologic
phenomena.
•
To understand blood disorders in the context of thestructural and functional features of the
hematopoietic-lymphoid system.
• To apply principles of basic sciences (biochemistry,
immunology, molecular biology and genetics) inunderstanding blood disorders
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References
• Hematology in Clinical Practice; Hillman RS et al(Paperback)
McGraw-Hill Professional; 4th edition (June 1, 2005)
• Essential Haematology; Hoffbrand V et al, Wiley-Blackwell;
5th edition (October 30, 2006)
• Harrison’s Textbook of Internal Medicine; Fauci et al (eds);
McGraw Hill, 17th edition, 2008
• Haematology An Illustrated Colour Text; Howard MR,
Hamilton PJ, 3rd Ed (2008) Churchchill Livingstone Elsevier
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INTRODUCTION
THE BLOOD
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Composition of the Blood
• Cellular element
– Red blood cells
– White blood cells
• Granulocytes
– Neutrophils
– Eosinophils
– Basophils
• Monocytes
•Lymphocytes
– Platelets
• Fluid elements
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RBC’s
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NEUTROPHILS
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LYMPHOCYTES
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MONOCYTES
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MONOCYTES
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BASOPHILS and EOSINOPHILS
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PLATELETS
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Composition of the Blood
• Fluid elements
– Water
– Electrolytes
–
Proteins• Albumin
• Immunoglobulins and
complements
• Transport proteins
– Transferrin
– Transcobalamin
– Haptoglobin Serum electrophoresis
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HEMATOPOIESIS
Origin and Development
of Blood Cells
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Hematopoiesis -
• Refers to the process of formation,
development, and differentiation of the
formed elements of blood.
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Hematopoietic Stem Cells (HSCs)
• Essential Attributes – Self-renewal
– Multilineage differentiation
– Reconstitution of hematopoietic system of a lethally
irradiated host• HSCs are rare, occurring at a frequency of 1 stem
cell per 10,000 to 100,000 bone marrow (BM)cells
• HCSs are quiescent (ie, noncycling), and only afraction enter into the cell cycle to proliferate anddifferentiate
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Embryology
• Hematopoietic cells originates frommesodermal tissue – migrates to theextraembryonic yolk sac – primitive
hematopoiesis composed of erythroid cellsand endothelial cells --- common precursor – hemangioblasts
• Definitive hematopoiesis ---- aortic-gonad-
mesonephros (AGM) ---- fetal liver and spleen---- bone marrow
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HEMATOPOIETIC STEM CELLS
Deri ation and Fate of HSCs
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Derivation and Fate of HSCs
Models of HSC self-renewal and
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Models of HSC self renewal and
differentiation
Hierarchical organization of hematopoiesis
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Hierarchical organization of hematopoiesis
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Stem and Progenitor Cells
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Stem and Progenitor CellsCharacteristic Stem Cell Progenitor Cell
Proliferative potential Tremendous More limitedRenewal On a population basis Probably none
Potential for differentiation All lineages Restricted
Differentiated characteristi Minimal-lineage negative Progressively increases
Functional characteristics Rapid directed motility and
expression of proteopodia;
homing
Less defined
Cycle status Dormant Cycling
Cytokine responsiveness Large number of cytokines
needed
Restricted
Cell o f origin Unknown Stem cell
Rhodamine and Hoechst
dyes stain
Stains dimly Bright
Adheres to marrow stroma Yes No or limited
Site of Hematopoiesis
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Figure 20-03
Copyright © 2005 Elsevier Inc. (USA) All rights reserved.
Site of Hematopoiesis
M S
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Marrow Structure
B M Mi i
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Bone Marrow Microenvironment
Marrow Microenvironment
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Marrow Microenvironment
Role of Marrow Microenvironment in
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Role of Marrow Microenvironment in
Hematopoiesis
• Provide a source of growth factors
• Important in the “trafficking” of immature
HSCs into and out of the marrow and the
blood. (Homing and Mobilization)
The Hematopoietic Hierarchy
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Figure 17-01
Copyright © 2005 Elsevier Inc. (USA) All rights reserved.
p y
Ch t i ti f HSC
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Characterization of HSCs
• Surface markers
• Dye exclusion
• In vitro culture
• Enumeration of HSCs by transplantation
models
S f HSC
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Sources of HSCs
• Bone marrow
• The peripheral circulation after mobilization
by chemotherapy and cytokines (mPB),
• Umbilical cord blood
H i i
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Hematopoiesis
Operational Components of
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p p
Hematopoiesis
• Anatomical structure of the marrow
• The stroma or marrow microenvironment
• Hematopoietic stem cells
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HEMATOPOIETIC STEM AND PROGENITOR CELLS
Characteristic Stem Cell Progenitor Cell
Proliferative potential Large More limited
Renewal Yes Probably none
Potential for differentiation All lymphohematopoieticlineages
Restricted
Differentiated
characteristics
Minimal-lineage negative Progressively increases to
lineage specific cell
Cycle status Dormant Cycling
Cytokine responsiveness Large number of cytokinesneeded for expression of
phenotype
Restricted
Cell of origin Unknown Stem cell
Adheres to marrow stroma Yes Limited to none
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The Humoral Regulation of
HematopoiesisCytokines
Characteristics of Cytokines
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Characteristics of Cytokines
• Are glycoproteins• Act on cell surface receptors
• Initiate complex second messenger and post-translational regulation
•May act on stem cells, progenitor cells, anddifferentiated cells of the same lineage
• May act on multiple different lineages
• Stimulate or inhibit proliferation, apoptosis,differentiation, or function
• Usually act on neoplastic counterpart of normal targetcell
Varying Tissue Origin of Cytokines
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Copyright ©2008 American Society of Hematology. Copyright restrictions may apply.
Metcalf, D. Blood 2008;111:485-491
No Caption FoundVarying Tissue Origin of Cytokines
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Copyright ©2008 American Society of Hematology. Copyright restrictions may apply.
Metcalf, D. Blood 2008;111:485-491
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Metcalf, D. Blood 2008;111:485-491
No Caption Found
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Metcalf, D. Blood 2008;111:485-491
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Cytokines with Hematopoietic
Activities
Colony stimulating factors
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Colony-stimulating factors
• Granulocyte colony-stimulating factor (G-CSF)• Granulocyte-macrophage colony-stimulating
factor (GM-CSF)
•Macrophage colony-stimulating factor (M-CSF)
• Interleukin-3 (IL-3)
• Erythropoietin (EPO)
•Thrombopoieitin (TPO)
• Interleukin-5 (IL-5)
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Figure 19-01
Copyright © 2005 Elsevier Inc. (USA) All rights reserved.
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HEMATOPOIETIC PROGENITORS ASSAYED IN VITRO
Cell Type Lineage Primary Cytokines
CFU-GEMM Neutrophil, erythroid, macrophage, T
cell, megakaryoctes
IL-3,GM-CSF, EPO
CFU-GM Neutrophil, monocytes-macrophages GM-CSF
CFU-M Monocyte-macrophage M-CSF (CSF-1)
CFU-G Neutrophil G-CSF
CFU-EOS Eosinophil IL-5, IL-3, GM-CSF
BFU-E Erythroid IL-3, KIT Ligand, GM-CSF,
IL-4, EPO
CFU-E Erythroid EPO
BFU-meg Megakaryocytes IL-3, GM-CSF,c-mpl ligand
CFU-meg Megakaryoctes IL-3, GM-CSF, c-mpl ligand,
IL-11, IL-6, LIF, G-CSF, KIT
ligand, IL-4
CFU-baso Mast cell, basophil IL-3, KIT ligand, GM-CSF
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Table 19-02
Copyright © 2005 Elsevier Inc. (USA) All rights reserved.
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The Bone Marrow
The Hematopoietic
Microenvironment
Cellular Components of the Marrow
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Microenvironment
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“THROUGHOUT THE CENTURIES
THERE WERE MEN (WOMEN)WHO TOOK FIRST STEPS
DOWN NEW ROADS
ARMED WITH NOTHING
BUT THEIR OWN VISION”
AYN RAND
THE FOUNTAINHEAD
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HEMATOPOIESIS
Part 2
Erythropoiesis
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Erythropoiesis
Erythron
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Erythron
Granulocytopoiesis
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Granulocytopoiesis