2 - Introduction To Hematopoiesis and RBC Production

HEM
311
Introduction to
Hematopoiesis and
RBC Production
Ma. Christy V. Gonzales, RMT, MPH
College of Medical Laboratory Science Our
Lady of Fatima University- Valenzuela
Hematology 1 (HEM 311)
HEM
311
Leaning
objectives At the end of this session, the students will be able to:
1. Explain the process of hematopoiesis.
2. List the cells found in bone marrow.
3. Describe the stem cell theory.
4. Explain the stages of maturation red blood cells.
2
Hematopoiesis
• Continuous and regulated process of cell production
• cell renewal
formation, development and
• cell proliferation
specialization of functional blood cell
• cell differentiation
• cell maturation
• BM
• Yolk sac AGM region

Hematopoiesis
Hematopoietic Development
1. Mesoblastic Phase
• 19th day of embryonic development after fertilization
• mesoderm to yolk sac
• central cavity of yolk sac
• surround cavity of yolk sac
• form blood vessels
• primitive but transient yolk sac erythroblasts
• important in early embryogenesis for hemoglobin production
• occurs intravascularly
• cells from mesoderm also migrate to AGM region
• HSCs Hematopoietic Stem Cells
2. Hepatic Phase
• 2 to 3 month of fetal development
nd rd
• 6 month
th
• 1-2 weeks after birth

• occurs extravascularly
• Production of megakaryocytes
• Thymus
• 1st fully developed organ in fetus
• Kidney & spleen
• Produce B cells
• Spleen
• Decline granulocytic production;
increases lymphocytic production
• Predominant Hb F and detectable
Hg A
3. Myeloid Phase
• Before the 5th month of fetal development
• BM cavity
• Medullary hematopoiesis
• HCSs and mesenchymal cells migrate into the core of the BM
• Mesenchymal cells
• Type of embryonic tissue
• Differentiate into structural elements that support developing
blood cells
3. Medullary Phase
• Myeloid-to-erythroid ratio 3:1 to 4:1
• End of the 24 weeks of gestation
• BM becomes the 1° of hematopoiesis
• Flat bones
• Tibia, pelvic area, sternum, vertebra
• Hb F and Hb A
• EPO
• G-CSF, GM-CSF
Blood cells
Hematopoietic stem cells (HSCs)
• foundation of the adult hematopoietic system
• embryo produces the first adult repopulating HSCs
Types of Human Stem Cells

1. Totipotential stem cells
2. Pluripotential stem cells
3. Multipotential stem cells
1. Totipotential stem cells
• presence: 1st few hours after an ovum is fertilized
• most versatile type of stem cell
• develop into any human cell type (from embryo into fetus)
2. Pluripotential stem cells

• presence : days after fertilization
• develop into any cell type, EXCEPT cannot develop into a fetus
3. Multipotential stem cells
• from pluripotent stem cells
• found in adults, but limited to specific types of cells to form tissues
• bone marrow stem cells can produce all types of blood cells, bone cartilage, and adipose (fat)
cells
• a fetus
Bone Marrow
• found within the cavities of all bones due to resorption of cartilage and endosteal bone
• trabeculae
• radiate out from the bone cortex into the central space
• provide structural support for the developing blood cells
• 2 forms:
• Yellow marrow
• normally inactive and composed mostly of fat tissue
• with undifferentiated mesenchymal cells and macrophages
• Red marrow
• normally active in the production of developing blood cells and their progenitors
Bone Marrow
• one of the body’s largest organs
• approx. 3.5% to 6% of total body weight
• ave. around 1,500 g in adults
• consists of hematopoietic cells
• erythroid, myeloid, lymphoid, and megakaryocyte
• fat (adipose) tissue
• osteoblasts and osteoclasts
• stroma
• hematopoietic cell colonies are compartmentalized
Bone Marrow
• Cords cross the walls of sinuses circulation
• First years of life: red and cellular

• 5 and 7 years of age: yellow marrow (retrogression)
• Active marrow restricted in:
• Proximal ends of long bones
• Vertebrae, ribs, sternum,
• skull bones, pelvis
• marrow cellularity
• ratio of red to yellow
• decreases with age
Bone Marrow
Stromal Cells
• endothelial cells, adipocytes macrophages and
lymphocytes, osteoblasts, osteoclasts, and reticular
adventitial cells
• endothelial cells
• broad, flat cells
• regulate the flow of particles entering and leaving
hematopoietic spaces in the vascular sinuses
• secrete cytokines
• adipocytes
• large cells with a single fat vacuole
• regulating the volume of active marrow
• secrete cytokines or growth factors
• macrophages
• phagocytosis
• secrete cytokines
Stromal Cells
• osteoblasts
• bone-forming cells
• osteoclasts
• bone-resorbing cells
• reticular adventitial cells
• secretes cytokines
• incomplete layer of
cells in vascular
sinuses
• form a supporting lattice for the developing hematopoietic
cells
• regulation of hematopoietic stem and progenitor cell survival and
differentiation
Red marrow
• compose of developing cells in extravascular cords
• located in spaces between vascular sinuses
• supported by trabeculae
• separated from the lumen of the vascular sinuses by
endothelial and reticular adventitial cells
Red marrow
• locations of developing cells:
1. Erythroblasts
• small clusters throughout the red marrow
• mature forms: adjacent to the outer surfaces of the vascular
sinuses
Red marrow
• locations of developing cells:
2. Megakaryocytes
• adjacent to the outer surfaces of the vascular sinuses
• release of platelets into the lumen of the sinus
3. Immature myeloid cells
• metamyelocyte
• deep within the cords
Bone Marrow
• Extramedullary hematopoiesis
• Abnormal
• When spleen, liver and lymph nodes revert back to produce immature blood cells
• enlargement of the spleen and liver
• Reasons:
• BM becomes dysfunctional
• BM is unable to meet the demands placed on it

Stem Cell Theory of Hematopoiesis
• All cells are derived from a pool of stem cells that are self-renewing
• Pluripotential & multipotential stem cells give rise to committed stem cells for each cell line
• Committed stem cells have receptors for specific growth factors
• Respond to stimulation by division & maturation (precursor cell stages) into end-stage cells
Hematopoiesis
Theories describing the origin of HSCs:
1. Monophyletic theory
• cells are derived from a single progenitor stem cell called a pluripotent hematopoietic stem cell
2. Polyphyletic theory
• each of the blood cell lineages is derived from its own unique stem cell
Theories describing the fate of HSCs:
1. Stochastic model of hematopoiesis

• random process whereby the HSC randomly commits to self-renewal or differentiation
2. Instructive model of hematopoiesis

• microenvironment in the bone marrow determines whether the HSC will self-renew or
differentiate
3 phases of HSC according to cell maturity:
• Primitive, multipotential/pluripotential cells
• most immature group capable of self-renewal and differentiation into all blood cell lines
• Intermediate cells
• consists of committed progenitor cells destined to develop into distinct cell lines
• Mature cells
• most developed group with specific functions
Stem Cell Theory
of
Hematopoiesis
Stem Cell Theory
of
Hematopoiesis
Regulators
Cytokines & Growth factors
• regulate the proliferation, differentiation, and maturation of hematopoietic precursor cells
Cytokines
• soluble proteins that have direct and indirect effects on hematopoietic cells
• include interleukins (ILs), lymphokines, monokines, interferons, chemokines, and colony-
stimulating factors (CSFs)
• stimulation (+; KIT ligand, FLT3 ligand, GM-CSF, IL-1, IL-3, IL-6, and IL-11) or inhibition
(-; transforming growth factor-beta, TNF-alpha, & interferons ) of production, differentiation, and
trafficking of mature blood cells and their precursors
Regulators
Growth factors
• regulating the proliferation and differentiation of HSCs
• regulating the survival and function of mature blood cells
• specific binding to receptors on the surface of target cells.
• Chromosome 7
• Gene for EPO
• Long arm of chromosome 5
• GM-CSF
• IL-3
• M-CSF
• Chromosome 17
• G-CSF
Regulators
Regulators
Colony-Stimulating Factors
• high specificity for their target cells
• active at low concentration
• names indicate the predominant cell lines that respond to their
presence
• G-CSF: granulocytic cell line
• GM-CSF: granulocytic-monocytic cell line
• GM-CSF + IL3: megakaryocyte cell line
Regulators
Interleukins
• protein molecules work in conjunction w/ hematopoietic growth factors
• to stimulate proliferation and differentiation of specific cell lines
• cytokines that act independently or in conjunction with other interleukins to encourage
hematopoietic growth
• cell signaling molecules
• first described as signals for communication between (inter—between) white blood cells (leuk— from
leukocytes)
Regulators
Regulators
Regulators
Regulators
Regulators
Marrow Differential
General Cell Characteristics
1. Overall cell size
• compared with the size of a mature erythrocyte
• erythrocytes and leukocytes
• megakaryocytes
2. Nuclear-cytoplasmic ratio
• amount of space occupied by the nucleus in relationship to the space
occupied by the cytoplasm
• size of nucleus generally decreases as a cell matures
• N:C ratio decreases
Features for Nuclear ID
1. chromatin pattern
2. nuclear shape
3. presence of nucleoli
Features for Cytoplasmic ID

4. staining color and intensity
5. granulation
6. shape
7. quantity of cytoplasm
8. vacuolization
9. inclusion bodies
1. chromatin pattern
• most distinctive nuclear feature of a cell in terms of maturity and cell type recognition
• Lymphocytes
• smooth or homogeneous pattern of chromatin throughout development
• Granulocytes
• fine to a highly clumped pattern
• Monocytes
• lacy pattern, which becomes finer as the cell matures
• Erythrocytes
• more clumped pattern as maturation progresses, until the extremely dense nucleus is lost from the
mature cell
2. nuclear shape
• round or oval and slightly folded nuclear shape
• Lymphocytes
• continue to have a round or oval nucleus
• Monocytes
• kidney bean–shaped nucleus, but folded or horseshoe shapes are common
• Neutrophils, eosinophils, and basophils
• segmented nuclei attached to one another by fine filaments with 2-5 depending on the cell
type.
• erythrocytes, leukocytes, and megakaryocytes
• all have nucleoli in the earliest cell stages
• as cells mature, nucleoli are usually not visible
• changes in appearance of nucleoli
rate of synthesis of ribosomal RNA
• number of nucleoli varies depending
on the cell type
• Lymphoblasts
• 1 or 2 nucleoli
• Myeloblasts
• 1 to 5 nucleoli
• Monoblasts
• 1 or 2 nucleoli but occasionally may have 3 or 4
• Erythroblasts
• not have any nucleoli or may have up to 2nucleoli that may stain darker than in other types of blast
cells
• Megakaryoblasts
• typically have 1 to 5 nucleoli
1. staining color and intensity
• in a Wright-stained blood smear
• vary with cell maturity and type
• from darker blue in younger cells to lighter blue or pink in mature cells
• Immature erythrocytes
• very distinctive dark-blue cytoplasm that becomes paler and gray looking as the cell
synthesizes hemoglobin
• Lymphocytes
• pale sky-blue cytoplasmic color
2. granulation
• presence, size, and color of granules are important in cellular identification
• no granules---nonspecific granulation---specific granulation
• blast forms of leukocytes and megakaryocytes
• Erythrocytes
• Granulocytes
• Variation
a. In size
b. In color
c. In the amount of granulation per cell
3. cytoplasmic shape
• useful in cellular identification
• Pseudopods
• mature monocytes and in some leukocyte blast forms
• Megakaryocyte
• develops a more irregular outline as the cell matures
• blast forms
• monocytes
• megakaryocytes
4. quantity of cytoplasm
• actual quantity of cytoplasm increases with age
• megakaryocyte
• extensive quantities of cytoplasm
• lymphocytes
• abnormalities frequently
5. vacuolization
• Monocytes
• having vacuoles throughout their life cycle and under normal conditions
• commonly seen in older cells and in abnormal conditions.
• Anticoagulants
• produce vacuoles as artifacts if the blood is stored for a longer-than-acceptable period
• Severe bacterial infections, viral infections and malignancies
• produce a remarkable number of vacuoles in various leukocyte types
6. inclusion bodies
• indicative of specific diseases
• Auer bodies or Auer rods
• myelocytic or monocytic blast forms
• erythrocytic inclusions and leukocytic inclusions
• Wright-stained blood smear and special stains
Erythropoiesis
• process of erythrocyte production

• differentiation from the HSC through the mature erythrocyte
• potential to differentiate into lymphoid or other hematopoietic cell types is
restricted
• Site:
Yolk sac---extramedullary organs---red bone marrow

Erythropoiesis
• mature erythrocyte
• biconcave disc with a central pallor
• Hemoglobin
• Respiratory protein
• Heme protein
• 120 days
• amino acids, iron, vitamin B12, vitamin B6, folic acid and the trace
minerals
• 200 billion erythrocytes = >20 mg of elemental iron
Erythropoiesis
Erythropoietin
• Site of production: Peritubular cells of kidney
• Liver – 10-15%
• Characteristics:
• Can cross the placenta
• 1st human hematopoietic growth factor
• Detected in phase
• EPO blood level α tissue oxygenation
• 20 mU/mL
• heme protein
• involved in the oxygen-sensing mechanism
Erythropoiesis
Erythropoietin
• predominant effect on the committed erythroid cells
• promote proliferation and differentiation of CFU-E
• stimulate the differentiation of a BFU-E
• prevents erythroid cell apoptosis
• reticulocyte
• Lipoxygenase
• Bcl-x
• interacts with IL-3, GMCSF, IL-1 and MEG-CSF
Erythropoiesis
Erythropoietin
• increase in the production of several types of RNA
• increase in DNA activity
• protein synthesis
• # of cells at each stage before the polychromatophilic erythroblast stage > at each preceding stage
• after polychromatophilic erythroblast stage, erythroid cells do not divide
• undergo specialized maturation
• increased erythrocyte production
• hemoglobin synthesis
Erythrocyte Maturation & Development
Early Cells
Pluripotent stem cells
myeloid stem cell
CFU-GEMM
BFU-E
CFU-E Mature erythrocyte

Early Cells
Pluripotent stem cells
myeloid stem cell
CFU-GEMM
BFU-E
CFU-E Mature erythrocyte

Hematopoiesis
1. Rubriblast
• 12 to 19 um
• N:C ratio is 4:1
• Nucleus
• Large, round and contains 0-2 nucleoli
• dark appearing has fine chromatin pattern
• Cytoplasm
• stains a distinctive blue color in Wright stain
• lacks granules
• most iron for hemoglobin synthesis is taken into the cell
2. Prorubricyte
• 12 to 17 um
• Nucleus
• chromatin becomes more clumped
• Cytoplasm
• stains a distinctive blue color in Wright stain
• no evidence of the pink color
3. Rubricyte
• 11 to 15 um
• Nucleus
• increasingly clumped
• Cytoplasm
• variable amounts of pink coloration mixed with basophilia
• muddy, light gray appearance
4. Metarubricyte
• 8 to 12 um
• Nucleus
• chromatin pattern is tightly condensed
• nucleus will be extruded from the cell
• Cytoplasm
• acidophilic
5. Reticulocyte
• BM = PC
• Reticular appearance caused by remaining RNA
• (+) Supravital stain
• Polychromatophilia
• Retics with high amount of RNA residual
• blue appearance in Wright’s stain
• 7 to 10 um
• Anuclear
• Mitochondria and ribosomes
6. Mature Erythrocyte
• 6-8 um
• Radioactice chromium ( Cr)
51
• Determine survivavility
• 120 days
• central pallor of 1 to 3 μm
References:
• Lotspeich-Steininger e.t al; Clinical hematology principles,
procedures, correlations, Lippincott Company, 1992
• Turgeon, Clinical Hematology: Theory and Procedures 5ht
ed., Lippincott Williams & Wilkins, 2012
• Keohane et. al, Rodak’s Hematology: Clincal Principles 6th
ed., Elsevier, 2020
• DOH Dept Circ 2017-0173 Schedule of EQAS Application
69
Thanks
!
Any questions?
You can find me at:
mvgonzales@fatima.edu.ph
HEM
311

2 - Introduction To Hematopoiesis and RBC Production

Uploaded by

Document Information

Original Title

Copyright

Available Formats

Share this document

Share or Embed Document

Sharing Options

Did you find this document useful?

Is this content inappropriate?

Copyright:

Available Formats

2 - Introduction To Hematopoiesis and RBC Production

Uploaded by

Copyright:

Available Formats

HEM

• Yolk sac AGM region

• 1-2 weeks after birth

Types of Human Stem Cells

2. Pluripotential stem cells

• First years of life: red and cellular

• BM is unable to meet the demands placed on it

1. Stochastic model of hematopoiesis

2. Instructive model of hematopoiesis

Features for Cytoplasmic ID

• process of erythrocyte production

Yolk sac---extramedullary organs---red bone marrow

myeloid stem cell

CFU-E Mature erythrocyte

myeloid stem cell

CFU-E Mature erythrocyte

You might also like