School of Medicine

Biomedical sciences (anatomy course unit)

Hematology system

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 Session plan
 Histology of the blood

 RBC

 WBC

 Platelets

 Development of the blood cells(hemopoiesis)

 Development of RBC

 Development of WBC

 Development of platelets

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Histology of the blood

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 Histology of Blood
Blood is fluid CT in which blood cells are
suspended in a liquid extracellular material
called plasma

A human adult has about 5 liters of blood,

which is roughly 7-8% of total body weight

A typical sample of blood is 45% formed

elements (blood cells) & 55% plasma by
volume

The formed elements include: erythrocytes

(red blood cells), leukocytes (white blood
cells) and platelets
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 Functions of Blood
1. Functions principally as a vehicle for the transport of gases (oxygen &
carbon dioxide), nutrients, metabolic waste products, cells, &
hormones:

o Provides tissues and cells with nutrients and oxygen

o Carries waste products such as carbon dioxide and nitrogenous

wastes away from the tissue

o Transports hormones to target tissues

o Carries drugs, elements and cells that protects the body from
infections and foreign bodies

2. A homeostatic role based on its thermoregulatory and acid-base

balance.
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Composition of whole blood

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 Plasma
 a protein-rich intercellular aqueous solution (pH 7.4) that imparts
blood its fluid properties.

 Contains substances of low or high molecular weight that make up 7%

of its volume

 Plasma proteins account for approximately 7% of the dissolved

components, with the remainder including nutrients, nitrogenous
waste products, hormones, and many inorganic ions collectively called
electrolytes

 Through the capillary walls, the low-molecular-weight components of

plasma are in equilibrium with the interstitial fluid of the tissues

 The composition of plasma is usually an indicator of the mean

composition of the extracellular fluids in tissues 7
 Composition of plasma
1. Albumin: is the most abundant plasma protein, is made in the liver
and serves primarily in maintaining the oncotic pressure of the blood.
2. α- and ß-globulins: made by liver and other cells, include; transferrin
and other transport factors; fibronectin; prothrombin; lipoproteins &
other proteins entering blood from tissues
3. γ-globulins: which are immunoglobulins (antibodies) secreted by
plasma cells in many locations.
4. Fibrinogen: the largest plasma protein, also made in the liver, which
during clotting polymerizes as insoluble cross-linked fibers (Fibrin)
which block blood loss from small vessels
5. Complement proteins: a system of factors important in inflammation
and destruction of microorganisms
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 The functions of plasma
 Maintaining optimal blood volume and pressure

 Supplying critical proteins for blood clotting and immunity

 A medium for exchange of vital minerals such as Na+, K+, Ca ++, &
phosphorus to maintain proper ionic balance

 Nutrients, such as glucose, proteins, lipids, and hormones are carried

from site of absorption or synthesis and distributed to various areas of
the body by blood plasma.

 Moreover, plasma transports nitrogenous wastes (such as urea, uric acid

& creatinine), excess salt & water to be removed from the blood by
the kidneys

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 Blood Cells
1. Erythrocytes (RBC)

2. Leukocytes (WBC)

3. Thrombocytes (Platelets)

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 Erythrocytes (RBCs)
 RBCs are terminally differentiated cells

 lack nucleus & other cytoplasmic organelles

such as mitochondria RER, ribosomes, etc.
but packed with the O2 carrying protein
hemoglobin

 Under normal conditions, these corpuscles

never leave the blood vessels

 The normal shape of RBC is rounded, flexible

biconcave disc

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 Erythrocytes…cont’d
• They are approximately 7.5µm in diameter,
2.6µm thick at the rim & 0.75µm thick at the
center

• Average volume is 90-95 cubic micrometers

• This biconcave shape provides a large

surface-to-volume ratio and facilitates gas
exchange

• The normal concentration of erythrocytes is

approximately 3.9–5.5 million per microliter
in women and 4.1–6 million per microliter in
men
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 Erythrocytes…cont’d
• Cytoplasm contains hemoglobin

• Hemoglobin consists of four polypeptide

chains.

• Each polypeptide chain is bounded to iron

containing heme group

• life span: 120 days (4 months)

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 Functions of erythrocytes
a. Transport carbon dioxide from the tissues to the lungs.

b. The presence of enzyme ‘carbonic anhydrase’ in erythrocytes makes it

possible for carbon dioxide to combine with water and thereby be
transported from the tissues to the lungs in the form of bicarbonate

c. Transport oxygen from the lungs to the tissues.

d. Erythrocytes contain hemoglobin in the cytoplasm which combines

rapidly with oxygen to form ‘oxyhemoglobin’

e. When this reaches the tissues, oxygen is released and the hemoglobin
that remains is called ‘reduced hemoglobin’

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 Erythrocytes…cont’d
 How the structure of RBCs is well adapted to their function?

1. The biconcave shape of a RBC increase its surface area through

which gaseous exchange takes place

2. The cell membrane is highly selective allowing the exchange of

gases

3. The highly elastic cell membrane allows the cell to change its
shape into almost any shape and to squeeze inside narrow
capillaries.

4. The absence of a nucleus and organoids allow more space for more
HGB inside the cell

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 Medical applications
 Poikilocytosis: an increase in abnormal red blood
cells of any shape that makes up 10% or more of the
total population

o Results from abnormal hemoglobin (called HbS)

o RBCs will be destroyed and phagocytosed by the

macrophages of the spleen, bone marrow, & liver

 Macrocytes: erythrocytes with diameters greater

than 9 µm are called

 Microcytes: erythrocytes those with diameters less

than 6 µm are called.

 Anisocytosis: the presence of a high percentage of

erythrocytes with great variations in size 16
 Medical applications…cont’d
 Sickle cell disease: Inherited alterations in hemoglobin molecules(HbS)
are responsible for several pathologic conditions, an example of which
is sickle cell disease

o caused by a mutation of one nucleotide (a point mutation) in the gene

for the chain of hemoglobin

o The triplet GAA (for glutamic acid) is changed to GUA, which specifies
valine

o The consequences of this single substitution are profound

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 Medical applications…cont’d
 Anemia: is pathologic condition characterized by blood concentrations
of hemoglobin below normal values.

• Anemia may be caused by:

o Loss of blood (hemorrhage)

o Insufficient production of erythrocytes by the bone marrow

o Production of erythrocytes with insufficient hemoglobin, usually

related to iron deficiency in the diet; or

o Accelerated destruction of blood cells

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 Medical applications…cont’d
 Rouleaux formation

• RBCs adhering in columns side by side like a pile of coins

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 Leukocytes
 They are colorless because they contain no hemoglobin, however, each
cell has a nucleus.

 In the bloodstream, they are spherical in shape and mobile.

 According to the type of cytoplasmic granules and the shape of their

nuclei, leukocytes are classified into;

A. Granular leukocytes: which contain specific granules and lobulated

nuclei):

a. Neutrophils 54-62%

b. Eosinophils 1-3%

c. Basophils 0-1%

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 Leukocytes…cont’d
B. Agranular leukocytes: which do not contain specific granules with
non-lobulated nuclei:

i. Lymphocytes 25-33%

ii. Monocytes 3-7%

 Total leukocytic count: is the total number of leukocytes per cubic

millimeter.

 In normal adult the ‘total leukocytic count’ is between 45,00-

11,000 per cubic mm

 Differential leukocytic count: is the percentage of each type of

leukocytes to the total number of leukocytes

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Leukocytes…cont’d

Granulocytes

Agranulocytes

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Leukocytes…cont’d

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 Functions of leukocytes
1. The role of leukocytes in general is essentially a protective one.

o They protect the body in a number of ways against the infectious

organisms

2. Moreover, leukocytes perform their main functions outside the blood

stream after they have entered loose CT.

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 Neutrophils
 Are the most numerous leukocytes in peripheral blood and,
as such, account for about 54-62% of the circulating
leukocytes.

 Leave circulation in large numbers in response to bacterial

infection and tissue injury in acute inflammations.

 Size: 12-15µm in diameter

 Their nucleus is multi-lobulated (2-5 lobules)(usually three)

in appearance, connected to each other by thin threads of
chromatin.

 In females the neutrophils are characterized by appearance

of inactive X chromosomes small separate one lobule of the
nucleus of neutrophil looks like ‘‘drumstick like’’
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 Neutrophils…cont’d
 it’s multilobulated, so termed as polymorphonuclear leukocytes.

 The specific fine granules in the cytoplasm stain faintly with neutral dyes.

 They also contain few non specific larger granules termed “azurophilic
granules’’ as they stain with methylene azure dye.

 At EM level:-

 The cytoplasm contains few organelles. It is full of small dark specific

granules which contain alkaline phosphatase & phagocytin (bactericidal
protein) which can destroy bacteria engulfed by neutrophils.

 The non specific granules are fewer, course and darker.

 These are the azurophilic granules which correspond to lysosomes.

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Neutrophils…cont’d

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Neutrophils…cont’d

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 Types of granules present in neutrophils
1. Azurophilic granules (primary granules): are primary lysosomes as
large, less numerous, dense vesicles and have a major role in both
killing and degrading engulfed microorganisms

 Contain proteases and antibacterial proteins

 Myeloperoxidase (MPO): which generates hypochlorite and other

agents toxic to bacteria

 Lysozyme: which degrades components of bacterial cell walls

 Defensins: small cysteine-rich proteins that bind and disrupt the

cell membranes of many types of bacteria & other microorganisms

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 Types of granules present in neutrophils…cont’d
2. Specific secondary granules: are smaller and less dense, stain faintly
pink, and have diverse functions, including
 contain antimicrobial and bactericidal peptides (i.e., lysozymes,
lactoferrins), various enzymes e.g. ECM-degrading enzymes such as
type IV collagenase, phospholipase, as well as complement activators.
 delivery of additional bactericidal proteins to the phagolysosomes
3. Tertiary granules: are of two types.
 One type contains phosphatases (enzymes that remove a phosphate
group from a substrate) and is sometimes called a phosphasome.
 other type contains metalloproteinases, such as gelatinases and
collagenases, which are thought to facilitate the migration of the
neutrophil through the connective tissue.
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 Functions of neutrophils
are the first line of defense against invading organisms especially bacteria.

Neutrophils (in blood stream) are attracted to bacteria (in CT) by chemical
substance liberated by the organism a process known as ‘‘Chemotaxis’’

Neutrophils squeeze their way b/n endothelial cells of blood capillaries or

venules.

Once neutrophils are outside the vessel in the CT they move toward
bacteria, which they rapidly phagocytose and destroy.

Neutrophils are the major constituents of tissue exudates and pus, and
thus they are called pus cells

Once the bacteria is engulfed within the cytoplasm of neutrophils, the

granules will act to destroy bacteria.

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 Eosinophils
 Characteristic feature of eosinophils

 Large cell size (12-17 µm in diameter)

 Constitute about 1-3% of total leukocytes

 a characteristic bilobed nucleus

 Contain many large refractile crystalloid eosinophilic

granules

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 Eosinophils…cont’d
 Types of granules present in eosinophils

1. Specific granules: abundant large, refractile specific granules(about

200 per cell) that are stained by eosin (acidophilic)

 contain lysosomal enzymes that inactivate histamine (histaminase)

and also aryl sulftase which neutralize action of slow reacting
substance

2. Azurophilic granules: lysosomes containing hydrolytic enzymes.

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Eosinophils…cont’d

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Eosinophils…cont’d

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 Ultrastructurally the eosinophilic specific granules
 are seen to be oval in shape, with many many having a flattened
crystalline core containing major basic protein, an arginine-rich factor
accounting for the granule's intense acidophilia

o This protein constitutes 50% of the total granule protein.

o The major basic protein, along with eosinophilic peroxidase, other

enzymes and toxins, act to kill parasitic worms or helminths worms
and protozoa.

 Eosinophils also phagocytose antigen-antibody complexes and modulate

inflammatory responses in many ways.

 They are an important source of the factors mediating allergic

reactions and asthma.
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 Ultrastructurally the eosinophilic specific granules

TEM (right) of a sectioned eosinophil clearly shows the unique specific

granules, as oval structures with disk-shaped electron-dense crystalline
cores (EG). These along with lysosomes & a few mitochondria (M) fill the
cytoplasm around the bilobed nucleus (N).
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 Function of eosinophils
 Cytotoxic effects on parasites such as helminthic worms.

 Eosinophils also phagocytose of Anti-allergic (antigen-antibody

complexes) and modulate inflammatory responses.

 They are an important source of the factors mediating allergic

reactions.

 What is meant by eosinophilia?

o Means increase in number of eosinophils more than 3% of total

leukocytes.

o It occurs in allergy, chronic inflammation and parasitic infection

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 Basophils
• Large size cells (12-15 µm in diameter)

• Nucleus is divided in two irregular lobes but the large specific granules
overlying the nucleus

• Are the least numerous less than 1% of the total leukocytes.

1. Azurophilic granules: are the lysosomes of basophils and contain a

variety of lysosomal acid hydrolases

2. specific granules: stain typically stain purple with the basic dye and
are fewer, larger, and more irregularly shaped than the granules of
other granulocytes

o contain glycosaminoglycans, leukotrienes, histamine and heparin

o Have surface receptors for immunoglobulin E (IgE)

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Basophils…cont’d

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Basophils…cont’d

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 Function of basophils
 Mediate inflammatory response

 They bind immunoglobulin E (IgE) in allergic reaction and leads to

release of their vasoactive substance (histamine)

 Play a role in hypersensitivity (anaphylactic shock) (i.e. vasodilatation)

 Are thought to supplement the function of mast cells in immediate

hypersensitivity reactions

 Basophilia; means Increase in number of basophils (more than 1%)

o Occurs in allergy reactions and liver cirrhosis

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 Lymphocytes
 are the primary functional cells of the immune system and also
actively involved in autoimmune diseases, inflammatory responses,
allergic reaction, tumor control and transplantation rejection/graft
rejection.
 reveal large, round, almost cell filling condensed nucleus; cytoplasm
forms only a thin rim around the nucleus.
 Smallest in size but are the 2nd most numerous (25-33% of leukocytes
in circulation),increased number are commonly seen during viral
infections.
 are the most common agranular leukocytes & are unique in their ability
to return from tissues back to the blood stream
 exist in two morphological forms: large & small lymphocytes
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Lymphocytes…cont’d

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Lymphocytes…cont’d

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 Lymphocytes…cont’d
 three groups of lymphocytes can be identified according to size: small,
medium, and large lymphocytes, ranging in diameter from 6 to 30µm.

 In the bloodstream, most lymphocytes are small or medium sized, 6 to

15µm in diameter.

 Large lymphocytes: are large granular lymphocytes, about up to 18 µm

in diameter

o Fewer in number, account only about 5% of lymphocytes & they are

believed to be NK cells

 Small and medium Lymphocytes: about 6-8 µm in diameter, include T

and B lymphocytes

o The majority more than 90% are small lymphocytes.

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 Lymphocytes…cont’d
 Three functionally distinct types of lymphocytes are present in the
body: T lymphocytes, B lymphocytes, and NK cells.

 T-lymphocytes (80%): have a long life span and are involved in

cellmediated immunity.

o arise in the bone marrow and proliferates in the thymus & activate
B- lymphocytes

 B-lymphocytes (15%): have variable life spans and are involved in the
production of circulating antibodies.

o Do not pass to the thymus

o Mature to plasma cells, which produce antibodies

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 Lymphocytes…cont’d
 NK cells: are programmed during their development to kill certain
virus-infected cells and some types of tumor cells.

 They also secrete an antiviral agent, interferon γ.

 NK cells are larger than B & T cells & have a kidney-shaped nucleus.

 NK cells have several large cytoplasmic azurophilic granule easily

seen by LM, they are also called large granular lymphocytes
(LGLs).

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Lymphocytes…cont’d

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 Lymphocytes…cont’d
B-Lymphocytes T-Lymphocytes
Percentage 15% of the circulating 80% of the circulating
lymphocytes lymphocytes
Development In the bone marrow In the bone marrow from
from (CFU-Ly B) (CFU-Ly T)
Maturation & In the bone marrow In the cortex and
Immuno- medulla of the thymus
competency
Plasma membrane Have Fc receptors and Have T-cell receptors
antibodies
Site in Peripheral Widely distributed in In thymus dependent
Lymphoid Organs lymph node, spleen zone of lymph node,
spleen, tonsil, peyer’s
patch, etc
Life Span 3 months May live for years
Function Humeral immunity Cell mediated immunity
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 Monocytes
 Normal range 3-7% of Total Leucocyte Count

 The large blood cells in size (12 to 20µm in

diameter)

 Nucleus is large eccentric, indented and kidney

shaped

 Cytoplasm is bluish-grey and has small number of

azurophilic granules (lysosomes), with occasional
vacuole like spaces.

 After crossing the walls of postcapillary venules,

monocytes differentiate into macrophages in
connective tissues, microglia in the CNS,
osteoclasts in bone, etc.
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Monocytes…cont’d

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 Monocytes…cont’d
 In the EM, nucleoli may be seen in the nucleus, and a few rough ER,
free polyribosomes, glycogen granules, and small mitochondria are
observed.

 horse-shoe shaped/kidney shaped nucleus

 contains more cytoplasm than does the lymphocyte

 Monocytosis means: Increase in number of monocytes

o They increases in chronic inflammation

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 Function of monocytes
1. They are highly phagocytic cells.

2. They are transformed to macrophages in tissues & at the site of

inflammation.

3. They concentrate the antigens and present them to the lymphocytes

and other immune cells.

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 Thrombocytes
 Platelets: are small, nonnucleated, rounded or
oval fragments of cells surrounded by a plasma
membrane.

 Promote blood clotting and help repair minor

tears or leaks in the walls of blood vessels,
preventing loss of blood.

 Normal values: 150,000- 400,000/mm3

 Size: 2-4 µm.

 Life span is about 10 days

 show few mitochondria & microtubules with

actin and myosin.
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 Platelets…cont’d
• In stained blood smears, each individual

platelet is appear in clumps, with;

o very lightly-stained peripheral zone, the

hyalomere

o darker-staining central zone containing

granules, called the granulomere

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 Platelets…cont’d
 Structurally, platelets may be divided into four zones based on
organization and function.
A. Peripheral zone: consists of the cell membrane covered by a thick
surface coat of glycocalyx.
– The glycocalyx consists of glycoproteins, GAGs, and several
coagulation factors adsorbed from the plasma.
B. Structural zone: comprises microtubules, actin fi laments, myosin,
and actin-binding proteins that form a network supporting the plasma
membrane.
C. organelle zone: occupies the center of the platelet & consists of
mitochondria, peroxisomes, glycogen particles & at least three types
of granules.
D. membrane zone consists of two types of membrane channels.
o Open canalicular system: connected to invaginations of the plasma
membrane, which may facilitate platelets' uptake of factors such as
fibrinogen and serotonin from plasma
o Dense tubular system: set of irregular tubular vesicles & derived
from the ER and stores Ca2+ ions.
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Platelets…cont’d

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 Platelets…cont’d
• Platelets contain three types of granules:

I. Alpha granules: contain fibrinogen and coagulation factors

II. Delta granules: contain ADP, ATP, and histamine; serotonin taken
up from plasma

III. Lambda granules: contain hydrolytic enzymes

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 Blood clotting
 Primary aggregation: Disruptions in the microvascular endothelium, allow
the platelet glycocalyx to adhere to collagen.

o Thus, a platelet plug is formed as a first step to stop bleeding

 Secondary aggregation: Platelets in the plug release a specific adhesive

glycoprotein and ADP, which induce further platelet aggregation

 Blood coagulation: During platelet aggregation, fibrinogen from plasma

giving rise to a fibrin to form a blood clot, or thrombus

 Clot retraction: The clot that initially bulges into the blood vessel lumen
contracts slightly because of the interaction of platelet actin and myosin

 Clot removal: the endothelium and surrounding tunic are restored by new
tissue, and the clot is then removed, mainly dissolved by the proteolytic
enzyme plasmin 60
Blood clotting…cont’d

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 Medical application
• Hemophilia A and B are clinically identical, differing only in the deficient
factor.

• Both are due to sex-linked recessive inherited disorders.

• Blood from hemophiliac patients does not coagulate normally: the blood
clotting time is prolonged.

• Persons with this disease bleed severely even after mild injuries, such as a
skin cut, and may bleed to death after more severe injuries.

• The blood plasma of patients with;

– hemophilia A is deficient in clotting factor VIII or contains a defective

factor VIII, one of the plasma proteins involved in fibrin generation;

– in hemophilia B, the defect is in factor IX.

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 Medical application…cont’d
• Generally, only males are affected by hemophilia A, because the
recessive gene to factor VIII is on the X chromosome.

• Females may have one defective X chromosome, but the other one is
usually normal.

• Females develop hemophilia only when they have the abnormal gene in
both X chromosomes, a rare event.

• However, women with a defective X chromosome may transmit the

disease to their male children.

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The end!

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