HEMATOPOIESIS: ERYTHROPOIESIS HEMATOLOGY 1 | SY 2020 - 2021

I. Introduction
Hematopoiesis

§ dynamic and complex production of blood cells

§ encompasses the overall interactions of renewal, proliferation, differentiation, maturation and death
§ occur in the organs of the reticuloendothelial system (RES)
o includes the bone marrow, spleen, liver, thymus and lymph nodes
o RES functions in hematopoiesis, phagocytosis and immune defense

II. Phases of Fetal Hematopoiesis

1. Mesoblastic
§ Chief site: Yolk sac
§ Primitive hematopoietic organ during fetal stage
§ Occurs intravascularly
§ Primitive erythroblasts – gives out hemoglobin to satisfy the oxygen needs of the fetus
§ Begin at the 19th day of embryonic development
§ Hemoglobin produced:
§ Gower-1: 2 zeta, 2 epsilon
§ Gower-2: 2 alpha, 2 epsilon
§ Portland: 2 zeta, 2 gamma

2. Hepatic
§ Chief site: Fetal Liver
§ begins at 5th – 7th gestational weeks
§ Beginning of definitive hematopoiesis
§ Occurs extravascularly
§ Appearance of lymphoid cells and granular leukocytes
§ Start of megakaryopoiesis
§ Hemoglobin produced:
§ Hb F: 2 alpha, 2 gamma

3. Medullary or Myeloid
§ Chief site: Red Bone Marrow
§ Begins prior to the fifth month of fetal development
§ During the myeloid phase, HSCs and mesenchymal cells migrate into the core of the bone
§ M:E ratio is 3:1 – 4:1 (adult)
§ Hemoglobin produced:
§ Hb A1: 2 alpha, 2 beta
§ Hb A2: 2 alpha, 2 delta
§ Hb F: 2 alpha, 2 gamma

PREPARED BY: JENINA CAMILLE G. BULLAGO, RMT, DTA, MSMT | TRINITY UNIVERSITY OF ASIA 1
 HEMATOPOIESIS: ERYTHROPOIESIS HEMATOLOGY 1 | SY 2020 - 2021

III. Sites of Hematopoiesis

1. Medullary Hematopoiesis
§ Blood cell production within the bone marrow. Begins in the fifth month of gestation and continues
throughout life

Bone Marrow
§ Tissue located within cavities of cortical bone
§ Types of marrow:
o Red – hematopoietically active
o Yellow – inactive (adipocytes)

Infant Red Active Marrow

5-7 years old Retrogression Reticulocyte

Adipose
Undiferrentiated mesenchymal
cells

Adult Red marrow = yellow marrow

§ Retrogression à active marrow is replaced by adipocytes resulting to restriction of active marrow in the sternum,
vertebrae, scapula, pelvis, ribs, skull and proximal portion of long bones
§ Newborn: 80-90% of bone marrow is active red marrow
§ Young adult (20 years old): 60% of bone marrow is active
§ Older adult (55 years old): 40% of bone marrow is active; 60% fats
§ Cellularity - ratio of marrow cells to fat (red marrow/yellow marrow)
o Normocellular - 30-70% hematopoietic cells
o Hypercellular - >70% hematopoietic cells
o Hypocellular - <30% hematopoietic cells
o Aplastic - few or no hematopoietic cell
§ Active Red Marrow Hematopoiesis à RS3VP2 (Ribs, skull, scapula, sternum, vertebrae, proximal end of long
bones and pelvis)

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HEMATOPOIESIS: ERYTHROPOIESIS HEMATOLOGY 1 | SY 2020 - 2021

2. Extramedullary Hematopoiesis
§ Blood cell production outside the bone marrow. Occurs when the bone marrow cannot meet body
requirements
§ Due to:
o Bone marrow is dysfunctional - aplastic anemia, infiltration of malignant cells, leukemia
o Bone marrow is unable to meet the body’s demands – hemolytic anemia

2.1 Liver
§ Major site of blood cell production during the second trimester of fetal development
§ Functions of hepatocytes (adult):
§ protein synthesis and degradation
§ coagulation factor synthesis
§ carbohydrate and lipid metabolism
§ drug and toxin clearance
§ iron recycling and storage
§ hemoglobin degradation
§ Kupffer cells
§ secrete mediators for protein synthesis
§ Removes senescent cells and foreign materials
§ Liver pathophysiology
§ Porphyrias
§ Severe hemolytic anemias and RBC dysplasias
§ Extramedullary hematopoietic production

2.2 Spleen
§ Largest lymphoid organ in the body
§ Vital but not essential for life
§ Functions:
§ Receives 350 mL of blood
§ Synthesizes Immunoglobulin M
§ Storage of 30% of total platelets
o Splenomegaly = decreased platelets in the circulation
o Splenectomy = increased platelets in the circulation

• Parts:
• Peritoneum - exterior
• Connective tissue capsule - interior
a. White pulp: lymphocytes, macrophages and dendritic cells
b. Red pulp: cords of Billroth - specialized macrophages that filters blood

§ Methods of removing senescent or abnormal RBCs:

§ Culling – cells are phagocytized then organelles are degraded
§ Pitting – remove inclusions or damaged surface membrane

2.3 Lymph nodes

§ bean-shaped structures located along the lymphatic capillaries that parallel, but are not part of, the
circulatory system

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 HEMATOPOIESIS: ERYTHROPOIESIS HEMATOLOGY 1 | SY 2020 - 2021

• Regions:

§ Cortex – cortical region – cortical nodules – germinal centers/B cell proliferation

§ Paracortex - T cells and macrophages

§ Medulla –B lymphocytes and plasma cells

• Functions:
§ Site of lymphocyte proliferation
§ Processing of Ig
§ Filter debris and bacteria
§ Lymph is the fluid portion of blood that escapes into connective tube (low protein ; absence of RBC)

§ Superficial lymph nodes – inguinal, axillary, cervical and supratorchlear

§ Deep lymph nodes - retroperitoneal
§ Adenitis – infection of the lymph nodes

2.4 Thymus
§ Populated by primitive lymphoid cells from yolk sac and liver
§ Retains the capability to produce T lymphocyte
§ Size related to age
§ Cortex: “waiting zone”, densely populated with progenitor T cells
§ Medulla: “holding zone”; mature T cells
§ T helper cells – CD4+
§ T cytotoxic cells – CD8+
§ T suppressor cells – CD8+

IV. Theories on Blood Cell Formation:

1. Monophyletic theory
§ Also known as “unitarian”
§ Most widely accepted theory
§ All blood cells are derived from a single progenitor stem cell (pluripotent hematopoietic stem cell)
2. Polyphyletic theory
§ Also known as “dualistic”
§ Each of the blood cell lineages is derived from its own unique stem cell

Models of Hematopoiesis

• Stochastic

– Randomly commits to self renewal or differentiation

• Instructive

– BM microenvironment determine if stem cell will self renew or differentiate

• Multilineage priming model

– Cells’ fate determined by internal and external factors: hematopoietic inductive microenvironment

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 HEMATOPOIESIS: ERYTHROPOIESIS HEMATOLOGY 1 | SY 2020 - 2021

Cytokines
§ Also known as hematopoietic growth factors
§ regulate the proliferation, differentiation, and maturation of hematopoietic precursor cells

o interleukins
o Lymphokines
o Monokines
o Interferons
o Chemokines
o CSF

Types of Stem Cells:

§ Totipotential stem cells à the most versatile type of stem cell, including development from embryo
§ Pluripotential stem cells à can develop into any cell type, except they cannot develop into a fetus.
§ Multipotential stem cells à found in adults, but they are limited to specific types of cells to form tissues

Types of Maturation:
§ Symmetric division à Both daughter cells may follow the path of differentiation, leaving the stem cell pool

§ Asymmetric division à one daughter cell may return to the stem cell pool and the other daughter cell may follow
the path of differentiation

Hematopoietic Stem Cells (HSCs):

§ Multipotent stem cells that give rise to all the blood cells

1. Myeloid (monocytes and macrophages, neutrophils, basophils, eosinophils, erythrocytes,

megakaryocytes/platelets)

2. Lymphoid (T-cells, B-cells, NK-cells)

§ Fates of HSC
1. Self renewal
2. Differentiation
3. Apoptosis - physiological cell death that can be induced by deprivation of growth factors or prevented by
growth-promoting cytokines

***Cellular senescence - cells that have lived their life span and will die of old age

*** Necrosis - accidental cell death by phagocytic cells and is associated with lethal physical damage

V. Erythropoiesis
A. Introduction
§ ___________ à ___________ à ___________
§ _________________________
o glycoprotein produced by the kidney and the liver that induces Hb synthesis and differentiation factor

Precursor cells
– Include all cells antecedent to the mature cells
– Cellular compartments of the hematopoietic precursors
a) HSC

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HEMATOPOIESIS: ERYTHROPOIESIS HEMATOLOGY 1 | SY 2020 - 2021

b) Progenitor – restrictive developmental potential, not morphologically recognizable

c) Maturing cells

B. Nomenclature of Erythrocytic Stages of Maturation

C. Criteria used in identification of Erythroid Precursors:

1. Over all diameter of cell decreases
2. N:C ratio lowers as the cell matures
3. Nuclear chromatin pattern becomes coarser, clumped and more
condensed as it matures
a. __________________—condensed nucleus, no
parachromatin visible
4. Nucleoli disappear
5. Cytoplasmic color change:
a. From blue to gray blue (Basophilia)—immature
b. To Salmon Pink (eosinophilia)—mature

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HEMATOPOIESIS: ERYTHROPOIESIS HEMATOLOGY 1 | SY 2020 - 2021

D. Erythroid Precursors
1. PRONORMOBLAST
§ Nucleus: round to oval, containing 1 - 2 nucleoli
§ Cytoplasm: dark blue
§ Location: BM
§ Cellular Activity: Globin production
§ Length of time in this Stage: 24 hours

2. BASOPHILIC NORMOBLAST
§ Nucleus: Round to slightly oval, chromatin slightly condensed, 0-1 nucleoli
§ Cytoplasm: deeper, richer blue
§ Location: BM
§ Cellular Activity: Hb synthesis
§ Length of time in this Stage: 24 hours

3. POLYCHROMATIC NORMOBLAST
§ Nucleus: Round, chromatin quite condensed, no nucleoli observed
§ Cytoplasm: Gray blue
§ Location: BM
§ Cellular Activity: Hb synthesis
§ Length of time in this Stage: 30 hours

4. ORTHOCHROMIC NORMOBLAST
§ Nucleus: Round, fully condensed chromatin, no nucleoli observed
§ Cytoplasm: Increase in salmon pink color
§ Location: BM
§ Cellular Activity: Hb synthesis
§ Length of time in this Stage: 48 hours

5. POLYCHROMATIC ERYTHROCYTE
§ Nucleus: Absent, no nucleoli, no chromatin
§ Cytoplasm: Slightly more blue/ purple than a mature RBC
§ Location: BM
§ Cellular Activity: Hb synthesis
§ Length of time in this Stage: 24-48 hours

ERYTHROCYTE
§ Nucleus: Absent, no nucleoli, no chromatin
§ Cytoplasm: Salmon pink with central pallor
§ Location: Peripheral blood
§ Cellular Activity: Oxygen delivery
§ Length of time in this Stage: 120 days

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 HEMATOPOIESIS: ERYTHROPOIESIS HEMATOLOGY 1 | SY 2020 - 2021

E. Erythrokinetics

§ Term describing the dynamic of RBC production and destruction

§ ________________ à collection of all stages of RBCs throughout the body; the developing precursors in
the BM and circulating RBCs

Erythropoietin
§ major stimulatory cytokine for RBCs
§ thermostable, nondialyzable, glycoprotein hormone with a molecular weight of 34 kD
§ Reference Interval:
§ Adults: 4.1-19.5 mU/mL
§ Infants aged 3 weeks to 2 months: 5-13 mU/mL
§ Children aged 3 months to 16 years: 9-28 mU/mL

§ Major Effects:
1. Early release of reticulocytes to the circulation
§ induces changes in the adventitial cell layer of the marrow/sinus barrier that increase the width of the
spaces for RBC egress into the sinus.
§ downregulates the expression of surface membrane receptors of cells to decrease adhesion to BM

2. Inhibit Apoptosis
o Removing an apoptosis induction signal
i. binds to its receptor on the CFU-E reduce production of Fas ligand Fas + FasL = apoptosis

o stimulate production of various anti-apoptotic molecules Bcl-XL (now called Bcl-2 like protein 1)

3. Reduced marrow transit time

o increased rate of cellular processes
§ stimulates the synthesis of RBC RNA
§ accelerated hemoglobin production

o decreased cell cycle times

§ cessation of cell division early entry to circulation
§ reduced length of time between mitoses

RBC Destruction
Macrophage-Mediated hemolysis (Extravascular)
Lack of mitochondria à dec. glycolytic process à dec. ATP à dec. ATP-dependent enzymes à dec. selective
permeability of cell membrane à inc intracellular water and sodium à cell becomes spherical à trapped in splenic
sieve à ingested by macrophage

Mechanical hemolysis (Intravascular/ Fragmentation)

Breaks in BVà trap RBCà rupture RBC à release of cell contents in the plasma

PREPARED BY: JENINA CAMILLE G. BULLAGO, RMT, DTA, MSMT | TRINITY UNIVERSITY OF ASIA 8