Hematology
from mesodermal cells lining the yolk sac;
Module 2
Introduction
Blood is a fluid connective tissue originating from the
embryonic mesenchyme. The regulated process of cell
production that includes cell renewal, proliferation,
differentiation, and maturation is called Hematopoiesis.
It is a continuous process resulting in the formation,
development, and specialization of all the functional
blood cells that are released from the bone marrow.
In this module, the mechanisms for blood cell
production will be presented. The study of normal cell
development is important as any deviation from this
normal cell development may lead to hematologic
disorders or abnormalities.
General Principles of Hematopoiesis
Hematopoiesis (or Hemopoiesis) is the process
of blood cell production, including renewal, proliferation,
differentiation, and maturation. It is a continuous and
regulated process that results in the formation,
development, and specialization of all functional blood
cells that are released from the bone marrow. These
mature blood cells develop from a common
hematopoietic stem cell. This stem cell is capable of both
self-renewal and directed differentiation into the
different cell lineages – ie. the hematopoietic stem cell
can proliferate and can give rise to any of the functional
blood cell lineages.
Hematopoiesis in humans is characterized by
selective distribution of embryonic stem cells in specific
sites that are rapidly changing during the course of
development. In healthy adults, hematopoiesis is
primarily confined to the bone marrow. During fetal
development, blood cell development begins in the (1)
yolk sac, that later progresses to the Aorta-Gonad-
Mesonephros (AGM) region (mesoblastic phase); (2)
transfers to the fetal liver (hepatic phase); and (3)
terminally, resides in the bone marrow (medullary
phase).
Mesoblastic Phase:
• Starts at the 19th day after fertilization
• Progenitor cells of mesenchymal origin relocate
to the yolk sac; give rise to Hematopoietic Stem
Cells (HSCs)
• Erythroblasts (immature red blood cells) come
remaining cells surrounding the cavity develop
into angioblasts and later on form the blood
vessels
• Yolk sac differs from other phases of
hematopoiesis in that yolk sac hematopoiesis
occurs intravascularly (within the blood vessels)
• Primitive erythroblasts are differentiated from
later erythroblasts in that primitive erythroblasts
never lose their nucleus. These erythroblasts are
found in ‘blood islands’ surrounding a
macrophage called Nurse cell.
• Primitive erythroblasts start to produce the
following hemoglobin’s: Portland hemoglobin,
Gower 1 hemoglobin, and Gower 2 hemoglobin
• Some cells of mesodermal origin also transfer to
the AGM region to develop into HSCs for
definitive hematopoiesis.
Hepatic Phase
• Begins at around 4-5 weeks after fertilization;
Peaks at third month of development
• The liver becomes the primary site of
hematopoiesis
• Characterized by recognizable clusters of
myeloid cells.
• Lymphoid cells begin to appear
• Megakaryopoiesis (development of platelet
precursors, the megakaryocytes) begins
• Sites of secondary hematopoiesis: Thymus
begins to produce T cells; Spleen and kidneys
produce B cells
• With detectable levels of HbF (fetal
hemoglobin), HbA / HbA1 (major adult
hemoglobin), and HbA2 (minor adult
hemoglobin)
• Activity remains until 1-2 weeks after birth
Medullary Phase
• Starts at the 5th month of development; cells of
various stages of maturation in all lineages are
seen
• Mesenchymal cells transfer to the skeletal
tissues and develop into HSCs
• Myeloid to erythroid (M: E) ratio reaches 3:1
(adult M:E ratio) at the 21st week
• Bone marrow becomes the major site of
hematopoiesis. Shortly after birth, the BM
remains as the only tissue capable of blood cell
 production. When the BM is in distress or is not o Secondary lymphoid Organs: sites of
functioning properly, secondary hematopoietic activation of lymphocytes - Spleen,
organs such as the liver and spleen revert to their Lymph Nodes, Mucosa-associated
hematopoietic function (extramedullary lymphoid tissue (MALT), Gut-associated
hematopoiesis) lymphoid tissue (GALT)
• Erythropoietin, G-CSF, and GM-CSF (Growth
Factors) reach detectable levels Bone Marrow
• Hemoglobins produced are HbA / HbA1, HbF,
and HbA2 (minor adult hemoglobin)

Sites of Hematopoiesis in the developing individual

Organs involved in hematopoiesis: • Composed of red marrow and yellow marrow;

red marrow is the hematopoietic tissue and the
1. Bone Marrow – primary site of hematopoiesis in yellow marrow is composed of adipose
an adult • In adults, red marrow is located in the sternum,
2. Liver – major site of hematopoiesis during the skull, vertebrae, scapulae, ribs, pelvic bones, and
hepatic period proximal ends of long bones
3. Spleen – secondary site of hematopoiesis during • Ratio of red marrow to yellow marrow in the
the hepatic period
developing individual:
4. Thymus – secondary lymphoid organ; involved in
o Before birth: 100% Red Marrow
the maturation of T cells
o At birth: 90:10
5. Lymph Nodes – secondary lymphoid organ; o At 19-20 y/o: 60:40
involved in production of lymphocytes, filtration o In adulthood: 50:50
and removal of old and damaged cells o At 65 y/o: 40:60
6. Bursa equivalent organ – in humans, the Bursa-
• Yellow marrow can revert to red marrow when
equivalent organ is the bone marrow. In the
there is increased demand for hematopoiesis,
Fabricius bird, the bursa is the site of maturation
such as in acute blood loss and hemolysis
of B cells.
• The bone marrow is the site of production and
7. Mononuclear Phagocyte System
maturation of myeloid cells – erythrocytes,
8. Kidneys – produce erythropoietin
megakaryocytes, neutrophils, eosinophils,
9. Stomach – produces intrinsic factor
basophils, and monocytes
10. Yolk sac – site of primitive erythropoiesis
• The bone marrow produces the lymphocytes.
The B cells mature in the bone marrow. The T
Adult Hematopoietic Tissue:
cells, however, mature in the Thymus. The
lymphocytes are activated in secondary
• Bone Marrow is the major site of hematopoiesis
lymphoid organs.
• Lymphoid development occurs in primary and
secondary lymphoid organs:
o Primary Lymphoid Organs: sites of
maturation of lymphocytes - Bone
Marrow and Thymus
Liver
• Plays a significant role in hematopoiesis during
fetal life (hepatic phase)
• Responsible for synthesis of most proteins and
vitamins that play a role in regulating hemostasis
• Responsible for conjugating bilirubin from
hemoglobin breakdown
• Responsible for detoxification of blood
• Site of protein synthesis and degradation
• Kupffer cells lining the canaliculi remove
senescent and damaged red blood cells from
circulation as they pass through the liver
Spleen
• Largest lymphoid organ in the body; secondary
site of hematopoiesis during hepatic phase
Functions of the spleen:
1. Culling: removal of senescent (old) red blood
cells from blood circulation by phagocytosis
2. Pitting: removal of inclusion bodies from the
surface of red blood cells (ex. Pappenheimer
bodies – accumulated iron; Howell-Jolly bodies
– DNA remnants; Heinz bodies – globin
remnants)
3. Immune defense – it is a secondary lymphoid
organ, serving as a site of activation of
lymphocytes (B and T cells)
4. Storage of platelets – the spleen sequesters 1/3
of platelets produced to serve as reservoir
Thymus
• primary lymphoid organ; secondary site of
hematopoiesis during hepatic phase
• Site of maturation of T cells
Lymph Nodes
• Secondary lymphoid organ
• Site of activation of lymphocytes
• Filters debris, particulate matter, and bacteria
from the lymph
• Serves as site of proliferation of lymphocytes
Mononuclear phagocyte system
• composed of the monocytes and macrophages
Functions:
1. Phagocytosis – removal of debris, particulate
matter, and foreign cells from the blood
(monocytes) and tissue (macrophages)
2. Antigen Presentation – Antigens from digested
foreign cells (bacteria) are presented to T cells
for activation of the adaptive immune system
3. Mitogen (substances that promote mitosis)
secretion
4. Secretion of hematopoietic growth factors
(substances that influence the maturation and
differentiation of blood cells)
Kidneys
• responsible for production of erythropoietin
(growth factor that drives maturation of RBC
precursors) in response to hypoxia.
Erythropoietin acts on erythroblasts in the bone
marrow to stimulate proliferation and
maturation, for eventual release into the
circulation
Stomach
• produces intrinsic factor. Intrinsic Factor is
necessary for absorption of Vit. B12 in the
intestines. Deficiency of IF Leads to deficiency in
Vit. B12 and would lead to pernicious anemia (a
type of Megaloblastic anemia)
Stem Cell Theory:
Stem cells are characterized by its ability for/to:
1. self-renewal;
2. give rise to differentiated progeny (ie. a
hematopoietic stem cell can differentiate into a
common myeloid stem cell Of common
lymphoid stem cell to later on give rise to mature
and functional blood cells);
3. reconstitute the hematopoietic system in a
lethally irradiated individual
Normal cell development depends the interaction of:
1. Pluripotent stem cell
2. Microenvironment
3. Hematopoietic Growth Factors
A pluripotent hematopoietic stem cell can be
stimulated to undergo one of three possible fates: self-
renewal, differentiation, or apoptosis. When the stem
cell divides, it gives rise to two identical daughter cells.
The daughter cells may likewise be stimulated to
undergo any of the three outcomes. A stem cell can also
be stimulated for differentiation - eg. An HSC can
differentiate into a common myeloid stem cell (common
myeloid progenitor) or a common lymphoid stem cell
(common lymphoid progenitor). The common myeloid
stem cell may differentiate into committed (lineage-
specific) precursor cells such as a Proerythroblast to
eventually give rise to mature erythrocytes or
Megakaryoblast to give rise to platelets. Differentiation
of stem cells and maturation of precursor cells occurs
under the influence of hematopoietic growth factors and
under optimal conditions of the microenvironment.
Cytokines and Growth Factors: a group of
glycoproteins that regulate the proliferation,
differentiation, and maturation of hematopoietic
precursor cells. Cytokines can either promote or inhibit
proliferation, differentiation, and maturation of blood
cells. Cytokines may also inhibit apoptosis (programmed
cell death), allowing cells to proliferate. Cytokines may
be Colony Stimulating Factors (CSF), early-acting
multilineage growth factors or interleukins.
The diagram above shows the derivation of
hematopoietic stem cells and the sites of action of
cytokines. From Rodak’s Hematology: Clinical Principles
and Applications (Keohane, EM, Smith, LJ, & Walenga,
JM) ©2016.
General changes undergone by hematopoietic cells as
they mature:
1. As the cells mature, they decrease in size.
2. As the cells mature, cytoplasmic staining
becomes less basophilic.
3. As the cells mature, Nucleus-to-Cytoplasm (N:C)
ratio decreases.
4. As the cells mature, nuclear chromatin becomes
denser.
5. As the cells mature, nucleoli start to disappear.
Erythropoiesis
Erythropoiesis refers to the production and
development of red blood cells. This process is largely
influenced by erythropoietin (EPO). Erythropoietin acts
on erythroblasts (normoblasts; red blood cell precursors)
in the bone marrow, promoting proliferation and
maturation.
From the CFU-GEMM (Colony-Forming Unit –
Granulocyte, Erythrocyte, Monocyte, Megakaryocyte;
Common Myeloid Stem Cell), the earliest identifiable
colony of RBCs arises, the BFU-E (Burst Forming Unit –
Erythroid). The BFU-E gives rise to CFU-E (Colony-
Forming Unit – Erythroid) under the influence of IL-3,
GM-CSF, Thrombopoietin (TPO), and KIT Ligand. The
CFUE, under the influence of EPO differentiates into the
erythroid precursor Pronormoblast. Further
differentiation and maturation is also under the
influence of EPO.
Under normal conditions, the kidneys produce a
small amount of erythropoietin. But under conditions of
stress decreasing oxygen supply (hypoxia) to the tissues,
the kidneys respond to the stimulus by increasing
secretion of erythropoietin. This increased EPO output
likewise increases RBC production and release from the
bone marrow.
Maturation series of erythroid cells:

Normoblastic Nomenclature Erythroblastic Nomenclature Rubriblastic Nomenclature

Pronormoblast Proerythroblast Rubriblast
Basophilic Normoblast Basophilic Erythroblast Prorubricyte
Polychromatic (Polychromatophilic) Polychromatic (Polychromatophilic) Rubricyte
Normoblast Erythroblast
Orthochromic Normoblast Orthochromic Erythroblast Metarubricyte
Polychromatic (Polychromatophilic) Polychromatic (Polychromatophilic) Polychromatic (Polychromatophilic)
Erythrocyte* Erythrocyte* Erythrocyte*
Erythrocyte Erythrocyte Erythrocyte
Pronormoblast / Rubriblast
• Diameter: 12-20 um
• N:C ratio: 8:1
• Nucleus with 1-2 nucleoli
• Basophilic cytoplasm
• Heme Production starts in the mitochondria
and globin synthesis in the ribosomes begins
• May undergo mitosis
Basophilic Normoblast / Prorubricyte
• Diameter: 10-15 um
• N:C ratio: 6:1
• parachromatin area of the nucleus becomes
larger; Staining is purple-red
• Nucleus has 0-1 nucleolus
• Basophilic cytoplasm (darker than cytoplasm of
PN)
• Hemoglobin synthesis starts
• May undergo mitosis
Polychromatic Normoblast / Rubricyte
• Diameter: 10-12 um
• N:C ratio: 4:1
• Nuclear chromatin strands denser
• No visible nucleoli
• Cytoplasm: combination of blue and pink
(murky graY blue on Wright-stained smear)
• Last stage of Mitosis
Orthochromic Normoblast / Metarubricyte
• Diameter: 8-10 um
• Pyknotic Nucleus (condensed) without nucleoli
• N:C ratio: 1:2
• Cytoplasm stains pink due to predominance of
Hgb
• Last stage with a nucleus
Reticulocyte
• Diameter: 8-10 um
• Anucleate
• Cytoplasm stains pink
• Reticulum (cytoplasmic RNA remnants) may be
stained with supravital stains (eg. New
Methylene Blue) but not with ordinary stains
• Present in BM (about 2 days) but later released
into the peripheral blood
• Last stage where hemoglobin synthesis occurs
• Circulates as reticulocyte for 1 day before it
becomes a mature erythrocyte
Erythrocyte
• Diameter: 7-8 um
• Biconcave shape
• Has no mitochondria
• On a Wright-stained smear, appears as salmon-
pink or red-staining cell with a central pallor
area
• Circulates in the blood for 120 days before
being removed by splenic or liver macrophages
Leukopoiesis
Leukopoiesis refers to the production, development, and
maturation of white blood cells. Granulocytes and
Monocytesdevelop from the common myeloid stem cell
(CFU-GEMM) while the lymphocytes develop from the
common lymphoid stem cell. Leukopoiesis is subdivided
into three categories:
1. Granulopoiesis: development and maturation of
Neutrophils, Eosinophils, and Basophils;
2. Monopoiesis: development and maturation of
Monocytes and macrophages; and
 3. Lymphopoiesis: development and maturation of
Lymphocytes

Granulopoiesis
The granulocytes – the neutrophil, the
eosinophil, and the basophil – have similar stages of
development. Although the stages of development are
similar, that is not to say that they develop from the
same precursor - ie. the myeloblast stage is a stage that
is common to all three cell lines, but the myeloblast is
already committed to a single cell line. But because the
stages of development are the same, development of
these three cell lines are often discussed as a group.
Growth Factors involved in differentiation and
maturation are GM-CSF and G-CSF

Stages of development of granulocytes:

It is important to note, again that each of the

three cell lines do not develop from the same
Myeloblast. The Neutrophil develops from its own
precursor, the Neutrophilic Myeloblast. Although for
practical purposes, because all three myeloblasts look
alike, they are simply referred to as ‘Myeloblast.’

Summary of Morphological Changes in Granulocyte Development (Romanowsky-stained)

Cell Cell Size Cytoplasm Shape of N/C Ratio Nuclear Nucleoli
(diameter) Nucleus Chromatin
Myeloblast 15-20 um Basophilic/ Round to 4:1 Fine 2-5
primary slightly oval
granules
Promyelocyte 15-21 um Basophilic 3:1 to 2:1 Slightly 2-3
coarser
Myelocyte 12-18 um Secondary Round to oval 1:1 Coarse -
granules w/ flattened
side
Metamyelocyte te 10-15 um Indented Decreased Coarse and -
clumped
Neutro. Band 9-15 um Lilac-pink C, S or Z Coarse and -
granules clumped
Neutrophil 9-15 um Lilac-pink 2-5 lobes Coarse and -
granules clumped
Eosinophil 9-15 um Reddish- 2 lobes Coarse and -
orange clumped
granules
Basophil 10-16 um Bluish-black 3-4 lobes Coarse and -
granules clumped
Myeloblast:
• earliest recognizable stage under the light
microscope

There are three types of myeloblasts:

• Type I Myeloblasts: no visible granules when
viewed under the light microscope
• Type II and III blasts: granular when observed
under the light microscope

Promyelocyte:
• stage of synthesis of primary granules (aka. Contents of Primary (Azurophilic) Granules:
non-specific granules; azurophilic granules) • Myeloperoxidase
• Acid-β-glycerophosphatase
Myelocyte: • Cathepsins
• Last stage capable of mitosis • Defensins
• Stage of synthesis of secondary (specific) • Elastase
granules – these granules are lineage-specific • Proteinase-3
• Charcot-Leyden Crystal Protein (Eosinophil only)
Metamyelocyte:
• stage which is most abundant in normal bone Contents of Neutrophil Secondary Granules:
marrow; stage of development of tertiary • β2-microglobulin
granules (forneutrophils only) • Collagenase
• Nucleus is slightly indented • Gelatinase
• Lactoferrin
Band: • Neutrophil gelatinase-associated lipocalin
• Stage of granulocytic development that first (NGAL)
appears in peripheral blood; granules are the • Transcobalamin
same granules of mature granulocytes; has a
deeply indented nucleus (C-, S-, Z-shaped) that Contents of Neutrophil Tertiary Granules:
may fold on itself • Gelatinase
• Collagenase
Mature (Segmented) Neutrophil: • Lysozyme
• has fine lilac to purple granules in the • Acetyltransferase
• β2-microglobulin
cytoplasm; nucleus has 2-5 segments (thus, also
known as polymorphonuclear neutrophil /
PMN) Contents of Eosinophil Secondary Granules:
• Major Basic Protein
Mature Eosinophil: • Eosinophil cationic protein
• Eosinophil-derived neurotoxin
• has coarse orange or pink granules in the
• Eosinophil peroxidase
cytoplasm that do not overlap with the nucleus;
• Lysozyme
nucleus is often bi-lobed
• Catalase
• β-glucoronidase
Mature Basophil:
• Cathepsin D
• has coarse bluish-black granules obscuring the
• IL-2, IL-4, IL-5, IL-6
view of the nucleus; nucleus is more often not
• GM-CSF
visible when viewed under the light microscope
Contents of Basophil Secondary Granules:
• Histamine
• Platelet-activating factor
• Leukotriene C4
• IL-4 and IL-13
 • Vascular endothelial growth factors A and B Monocyte 15-20um in diameter;
• Chondroitin sulfate (eg. Heparan sulfate) with blue-gray cytoplasm
(often described as
Monopoiesis having ground glass
Development and maturation of monocytes. Monocytes appearance); with a
are precursors of macrophages. The major cytokine kidney bean-shaped
responsible for monopoiesis is the M-CSF. nucleus (described as
having brain-like
Summary of morphological changes in monocyte convolutions); it is the
development only stage found in the
Monoblast 12-20um in diameter; peripheral blood; it is the
with basophilic largest cell in circulation
cytoplasm; high N:C ratio Macrophage 15-85 um in diameter;
(3:1 to 4:1); with 1-2 with a round to reniform
nucleoli; similar in nucleus; found in
appearance to the peripheral tissues
Myeloblast
Promonocyte 14-18um in diameter; Lymphopoiesis:
with blue-gray • Production, Development, and Maturation of
cytoplasm; N:C ratio is Lymphocytes
2:1 to 3:1; with 0-1 • Lymphocytes are divided into three major
nucleolus; last stage groups: T cells, B cells, and natural killer (NK
capable of mitosis; cells)
confined to the bone
marrow
Summary of morphological changes occurring in
lymphocyte development
Cell Cell Size Cytoplasm Shape of Nucleus N/C Ratio Nucleoli
(diameter)
Lymphoblast 10-18 um Moderate to Round to oval 4:1 1-2
dark blue
Prolymphocyte 10-18 um Moderate to Round to oval 4:1 1-2
dark blue
Lymphocyte S: 8-10um M: 10- Sky blue Round and
12 um L: 12-16 compact
um
Plasma cell 8-20 um Eccentric

then transfer to secondary lymphoid organs for

In lymphocyte development, lymphoblasts are
produced in the bone marrow. The development of B
cells and T cells differ in their location. For B cells,
lymphoblasts differentiate into pro-B cells, which later
become pro-B cells and immature B cells within the Bone
Marrow. Immature B cells leave the Bone Marrow and
relocate into secondary lymphoid organs where they
may be activated. Activated B cells may undergo mitosis,
become memory B cells, or become antibody-producing
plasma cells.
T cell development on the other hand, occurs in
the Thymus. Lymphoblasts in the Bone Marrow relocate
to the Thymus where they differentiate into pro-T, pre-
T, and later on immature T cells. These immature T cells
activation. Activated T cells may either become effector
T cells or memory T cells. Activation of T cells causes what
are seen in peripheral blood as medium or large
lymphocytes
MODULE 3
Introduction:
Red blood cells circulate in the blood for 120
days performing their function of oxygen delivery from
the lungs to the peripheral tissues. Essential to this
function of oxygen delivery is normal RBC morphology.
When Red Blood Cells have abnormal morphology or
become senescent (old), they are marked for elimination
from the circulation. In this module, student will be given
a more detailed study into normal and abnormal red
blood cell morphology, as well as the processes that are
essential to normal RBC function.
Red blood cells with abnormal morphology will
be discussed together with corresponding clinical
correlations of each abnormal morphology.
RBC Structure and Function:
The erythrocytes (red blood cells; RBCs) are the
most numerous cells in the blood with an average of 5
million RBCs per microliter and function for oxygen
delivery from the lungs to the peripheral tissues (ie. from
areas of high oxygen tension to areas of low oxygen
tension). The RBC is anucleate and biconcave with an
average volume of 90 fL. The cytoplasm has abundant
hemoglobin essential for the performance of RBC
function.
RBCs are produced through normoblastic
maturation in the bone marrow, as discussed in Lesson
3. The nucleus. While present in developing normoblasts,
is extruded before the developing red blood cell leaves
the bone marrow to enter the circulation. Free
ribosomes and mitochondria likewise disappear from the
RBC shortly after its release from the bone marrow.
Without the mitochondria for energy production
through oxidative processes, ATP is produced in the
cytoplasm through anaerobic glycolysis to provide for
the cell’s energy needs. Eventually, oxidative processes
limiting the RBC’s lifespan to 120 days. Thereafter, the
cell’s reusable components – globin, iron, phospholipids,
and proteins – are disassembled and recycled.
RBC Metabolism
Lacking mitochondria, the cell relies on
anaerobic glycolysis for energy production. RBC
metabolism occurs through four pathways: (1) the
Embden-Meyerhof Pathway and the glucose diversion
pathways that branch off from it: (2) Hexose
Monophosphate Shunt, (3) Methemoglobin Reductase
Pathway, and (4) Rapoport-Leubering Pathway.
Embden Meyerhof Pathway:
• Performs 90% of glycolytic reactions to provide
energy for the cell
• 1 molecule of Glucose enters the pathway and is
converted into pyruvate and lactate through a
series of biochemical reactions with a net
production of 2 ATP
• The energy produced is required to keep K+ ions
inside the RBC and keep Na+ ions out
• Energy is also used to maintain membrane
flexibility
Hexose Monophosphate Shunt
• Aka. Pentose Phosphate Pathway
• Produces reduced glutathione; prevents
oxidative denaturation of hemoglobin (globin
portion)
• Oxidative and aerobic pathway
• Functionally dependent on Glucose-6-Phosphate
Dehydrogenase (G6PD)
Methemoglobin Reductase Pathway
• Maintains iron in the hemoglobin molecule in
the ferrous (Fe2+) state by the action of
Methemoglobin cytochrome C reductase
Rapoport-Leubering Pathway
• Produces 2,3-Diphosphoglycerate (2,3-
Bi(s)phosphoglycerate; 2,3-DPG; 2,3-BPG)
• Regulates the hemoglobin’s affinity for oxygen
Erythrocyte Function:
1. Delivery of gases: delivers oxygen to peripheral
tissues and to a lesser extent, delivers carbon
dioxide to the lungs for excretion (carbon dioxide
is soluble in plasma; oxygen is not)
2. Maintain pH homeostasis: hemoglobin serves as
a minor buffer system in the blood by accepting
or releasing carbon dioxide
Erythrocyte development:
The red blood cell circulates in the blood for 120
days. During this period, the red blood cells decreasesin
ATP, glucose, and sialic acid while increasing in calcium.
These changes all contribute to the diminishing function
of red blood cells. Thus after 120 days, the cells are
eliminated from circulation.
Mechanisms of Erythrocyte Destruction:
1. Erythrophagocytosis (physiologic or
pathologic): Physiologic destruction of RBC
occurs as a result of splenic macrophages or
Kupffer cells of the liver phagocytosing
senescent red blood cells. Pathologic
erythrophagocytosis occurs abnormally such as
in Hemolytic Anemias
2. Fragmentation: RBCs fragment as a result of
mechanical or traumatic stress such as in the
presence of clots in the blood vessels.
3. Osmotic lysis: occurs as a result of red blood cells
being exposed to hypotonic solutions (ie. solute
concentration is less in the external solution
compared to the intracellular fluid) causing
influx of water molecules into the RBC, causing it
to swell and lyse. Spherocytes are extremely
vulnerable to osmotic lysis.
4. Hemoglobin denaturation
RBC structure
• 6-8 um in diameter; 1.5-2.5 um thick (around 2.5
um at the thickest portion and 1.5 um at the
central pallor region); 80-100 fL in volume
• Has a central pallor area representing the area
previously occupied by its nucleus.
• Buff or reddish with Romanowsky stains
• With and increased surface-volume ratio (it is
flatter than it is round)
Membrane Proteins:
• Maintains the shape and deformability of the
cell; the red blood cell has to be able to change
its shape in order to fit into the smallest of the
capillaries
• Composed of receptors, antigens, enzymes
• Lipid membrane attached to protein membrane
skeleton with about 10-12 major proteins.
• Integral proteins: penetrates the lipid bilayer;
Includes glycophorins; High sialic acid residues
(responsible for slight electronegative charge of
the RBC membrane
o Band3: inorganic anion transport
protein; facilitates Cl ion transport; site
of Ii Blood Group antigens
o PAS-1/PAS-2: glycophorins; may serve
as receptors for microorganisms; sites of
MN antigens and Gerbich Blood Group
antigens
o Band 4.5: glucose transporter; site of
ABH antigens
• Peripheral proteins: membrane cytoskeleton;
maintains shape and deformability of the cell
o Bands 1 and 2 (spectrin): 25-30% of
membrane proteins; primary
cytoskeletal proteins; responsible for
elasticity of the cell, together with actin
and protein 4.1
o Band 5 (actin): 4.5%
• Enzymes
o Carbonic anhydrase
o Methemoglobin reductase
o Catalase
o G-6-PD
o Pyruvate Kinase
o Adenosine deaminase
o Aldolase
o LDH
o Glutathione phosphorylase
o Nucleoside phosphorylase
o Acetylcholinesterase
I LOVE YOU SO MUCH AND I WILL ALWAYS BE HERE FOR
YOU NO MATTER HOW HARD THE FIGHT AND
CHALLENGES WILL BE I WILL ALWAYS CARRY AND
SUPPORT YOU ALL THE WAY. YOU WILL NOT BE ALONE
ANYMORE FOR IN THIS JOURNEY YOU HAVE ME AND, ON
THIS ADVENTURE, I AM WITH YOU, READY TO CATCH
YOU IF YOU FALL. LET” S GO AND KEEP PUSHING
FORWARD, I LOVE YOU SO MUCH.
-SEAN MELNOR P. LOSBAÑES