HAEMATOLOGY 1

PREPARED BY
TURYATUNGA BEN.
Granulopoesis

 Granulopoiesis is the production of neutrophils, eosinophils, and basophils.

 Granulocytic and monocytic cells are sometimes collectively referenced as myeloid
cells.
 However, the term myeloid and the prefix myelo- can be confusing because they have
other meanings; they may reference the bone marrow, all nonlymphoid hemic cells
(erythrocytes, leukocytes, and megakaryocytes), only granulocytes, or the spinal cord
Images
CONT…

 Production of the granulocytes (Myeloid stem cells Eosinophils, Neutrophils,

Basophils).
 Myelopoiesis (granulopoiesis) involves a series of maturational steps leading to the
formation of granulocytes, including neutrophils, eosinophils, and basophils.
 In the bone marrow, there is typically a spectrum of myeloid differentiation with
neutrophils and neutrophil precursors predominating. Because maturation is best
appreciated on a high-quality Wright–Giemsa-stained aspirate smear, the rest of the
section refers to this preparation.
 Myeloid blasts are infrequent (<2% of white blood cells) and vary in diameter from 15
to 20 µm, with a high nuclear:cytoplasmic ratio and scant pale blue agranular
cytoplasm.
Cont….

 The earliest granulocytic precursor identifiable by routine light microscopy is

the myeloblast, which undergoes maturational division over 5 days to produce 16 to 32
progeny cells.
 These granulocytic precursors are conceptually divided into those stages that can divide,
including myeloblasts, promyelocytes, and myelocytes (proliferation pool), and those that
cannot, including metamyelocytes, and band and segmented forms (maturation pool).
 Within the neutrophil maturation pool is a subpool, termed the storage pool, which
consists of a reserve of fully mature neutrophils.
 The size of the storage pool varies by species; it is large in the dog, but small in ruminants.
 In homeostasis mostly mature segmented granulocytes are released from the marrow into
the blood.
Cont….

 Myeloid blasts differentiate into promyelocytes, which have basophilic cytoplasm

with distinct magenta-colored primary cytoplasmic granules.
 Promyelocytes differentiate into myelocytes, which have slightly oval nuclei and pale
pink cytoplasm containing small, light ill-defined pink-reddish granules.
 Both promyelocytes and myelocytes may have a perinuclear halo.
 Myelocytes differentiate into metamyelocytes, which have a kidney-shaped nucleus
and granular cytoplasm.
 Metamyelocytes differentiate into bands with horseshoe-shaped nuclei. Ultimately,
these bands develop segmented nuclei and exit to the peripheral blood as mature
neutrophils.
 Eosinophils and basophils go through parallel stages of differentiation, but these cell
types are significantly less frequent in the marrow.
Cont…

 The main purpose of granulocytes is to migrate to sites of tissue inflammation and

function in host defense
 Briefly, these cells have key immunologic functions, including phagocytosis and
microbicidal activity (neutrophils and monocyte-derived macrophages), parasiticidal
activity and participation in allergic reactions (eosinophils and basophils), antigen
processing and presentation, and cytokine production (macrophages).
 Neutrophils are the predominant leukocyte type in blood of most domestic species
Regulation of granulopoiesis.

 G-CSF is a potent cytokine that influences the proliferation, survival, maturation, and
functional activation of cells from the neutrophil-granulocyte lineage
 However, in conditions of stress, G-CSF levels can rise to 20 times baseline levels,
thereby resulting in a rapid increase in circulating neutrophils.
 G-CSF may regulate this increased granulopoiesis by increasing the mitotic pool at the
promyelocyte and myelocyte stages and shortening neutrophil transit time in bone
marrow.
Stages of granulopoiesis.

• Pluripotential hemopoietic stem cell.

• Myeloblast.
• Promyelocyte.
• Eosino/neutro/basophilic myelocyte.
• Metamyelocyte.
• Band cell (Stab cell)
• Granulocytes (Eosino/neutro/basophil)
Myeloblast

 The earliest recognizable myeloid cell.

 Measuring (10- 20m)diameter with a large round to
oval nucleus/basophilic staining.
 It has a fine diffuse immature chromatin (without
clumping)
 A prominent nucleolus.
Promyelocyte

 Promyelocyte is the second stage of the Myeloblast development process.

 The promyelocyte is larger than the myeloblast.
 It has a diameter of 12-25µm and is the largest cell type in the myeloid
series.
 It has a prominent nucleus, and the nucleus is placed slightly intended in
the cytoplasm.
 Nuclear: Reddish-blue, Round, Coarse chromatin,
 Usually a nucleolus
 Cytoplasm: Bluish, Small, dark azurophilic granules
 larger than myeloblast
Cont….

 The cytoplasm of the promyelocyte is

granulated, and these granules are termed as
the primary azurophilic granules. Since the
promyelocyte is not differentiated
Myelocytes

 Are slightly smaller than Promyelocytes

 Measuring (10-18m) in diameter.
 With eccentric round-oval nuclei
 The chromatin is fine, but shows evidence of
condensation.
 Nucleoli may be seen in early stages but not in the
late myelocyte.
 Primary azurophilic granules are still present
Cont…

 Myelocytes belong to the third stage of granulopoiesis and are widely

studied as these are differentiated cells. Myelocytes is of three different
types such as, neutrophilic, eosinophilic and basophilic.
 Upon staining, the granules of the three different types of granulocytes
appear in three different colors.
 Granules of neutrophils – lilac
 Granules of eosinophils – orange-red
 Granules of basophils – purple
Cont…

 The structure of the myelocyte is similar to that of the promyelocyte but

has a reduced diameter.
 The cell diameter of the myelocyte is around 10- 20 µm.
 The myelocyte nucleus is an eccentric nucleus. The nucleus is oval or round
in shape, and one end is flattened.
 Nucleoli and chromatin structures are not very prominent and can only be
viewed under the electron microscope.
 The myelocyte is capable of cell division, and the proliferation of the
myeloid lineage cells stop at the stage of the myelocyte.
The Similarities Between Promyelocyte and
Myelocyte.

• Both are derived from the myeloid lineage.

• The precursor of both cells is the Myeloblast.
• Both cells participate in the process of granulopoiesis and result in the
development of granulocytes.
• Both cells are nucleated.
• Both cells are granulated.
• Both cells undergo cell division.
• Both cells have structures such as endoplasmic reticulum and Golgi
• Both cells can be stained to be observed under the light microscope or the
electron microscope
The Difference Between Promyelocyte and Myelocyte

Promyelocyte vs Myelocyte

Myelocyte is the third stage of myeloblast development which

Promyelocyte is the second stage of myeloblast development,
can be differentiated into eosinophils, basophils, and
and it is the largest cell type of the myeloid lineage.
neutrophils.

Size

Promyelocyte cell size ranges from 12 to 25 µm. Myelocyte cell size ranges from 10 to 20 µm.

Shape of the Nucleus

In myelocyte,the nucleus is an eccentric nucleus that is round

Nucleus in indented in promyelocyte.
or oval shaped.

Nucleoli and Chromatin Condensation

Prominent and seen under the light microscope in Not prominent, sparsely identified under the electron
promyelocytes. microscope in myelocytes.

Number of Granules

A high number of primary granules in the cytoplasm and in A low number of primary granules and secondary granules can
endoplasmic reticulum can be seen in promyelocytes. be seen in myelocytes.
Metamyelocyte
 In the metamyelocyte stage, the cytoplasm and nucleus continue to decrease in size.
 The cytoplasm achieves full secondary granule content. The chromatin becomes more dense, knotted, and
compact, while the nucleus begins to indent and acquire the familiar "kidney bean shape.
 By the end of the stage, the cell will be similar in size to a mature neutrophil with similarly cytoplasmic
granularity.
 Metamyelocyte [ neutrophilic and eosinophilic ] (10-18 µm): Note the indented nucleus (kidney
bean); light blue-purple and granular chromatin; no nucleoli are present; moderate clear pink
cytoplasm; specific granules are obvious. Basophilic metamyelocytes are very rare
Metamyelocyte images
BANDS/
stab(Neutrophil,Eosinophil,Basophil)
 The functional maturation of metamyelocytes results in the development of
band cells, which are usually slightly larger than mature neutrophils and
have a 15-µm diameter, a horseshoe-shaped nucleus, and a moderate to
abundant supply of specific granules.
 It is characterized by having a curved but not lobular nucleus.
Cont….

 A band neutrophil has moderate to abundant cytoplasm staining pale blue to pink.
Present within the cytoplasm are fine lilac to pink granules. These specific granules,
called neutrophilic granules, are small and somewhat resemble pencil stippling.
 The band neutrophil has a non-segmented nucleus. Instead, the nucleus shows an
indented shape which appears similar to a horseshoe. The nucleus of a band neutrophil
is typically U-shaped. Frequently, the nucleus of a band appears folded or twisted, thus
making identification a bit more difficult. The nucleus stains a deep purplish-blue color,
and the nuclear chromatin appears condensed, coarse, and clumped.
 Band neutrophils are also referred to as stabs or simply as bands. The diameter of a
band is approximately 9-16 microns, and its nuclear to cytoplasmic (N:C) ratio is
approximately 1:2.
 Band neutrophils are an intermediary step prior to the complete maturation of
segmented neutrophils.
 Polymorphonuclear neutrophils are initially released from the bone marrow as
band cells, as the immature neutrophils become activated or exposed to
pathogens, their nucleus will take on a segmented appearance.
 An increase in the number of these immature neutrophils in circulation can be
indicative of a infection for which they are being called to fight against, or some
inflammatory process.
 The increase of band cells in the circulation is called bandemia and is a "left
shift" process.
Band image
White cell lineage.
Cont….
White cells(granulocytes)

 Granulocytes: Mature granulocytes are plentiful; distinguished by the lobulated

nucleus, pink abundant cytoplasm, and specific granules.
Identify neutrophils , eosinophils ,and basophils (rare).
 Granulocytes are a type of white blood cell that has small granules. These granules contain
proteins. The specific types of granulocytes are neutrophils, eosinophils, and basophils.
 Granulocytes, specifically neutrophils, help the body fight bacterial infections.
 The number of granulocytes in the body usually increases when there is a serious infection.
 People with a lower number of granulocytes are more likely to develop bad infections more
often.
 Granulocytes are sometimes called granular leukocytes, polymorphonuclear leukocytes
or PMN.
The function of granulocytes

Granulocytes work together to rid your body of infection or allergens. Each type of
granulocyte has its own combination of chemicals and enzymes in its granules. As a
result, each type has a different function:
• Neutrophils: The most common type of granulocyte, neutrophils attack bacteria.
Each neutrophil cell can consume up to 20 bacteria in its lifetime.
• Eosinophils: These granulocytes are present in almost all immune responses, most
notably allergies. However, they also fight off parasites.
• Basophils: These granulocytes primarily combat allergic reactions. They release
histamine (which escorts allergens out of your body) and the blood thinner heparin
(which prevents clotting).
Functions of white blood cells.

• Neutrophils: Help protect your body from infections by killing bacteria, fungi and
foreign debris.
• Lymphocytes: Consist of T cells, natural killer cells and B cells to protect against viral
infections and produce proteins to help you fight infection (antibodies).
• Eosinophils: Identify and destroy parasites, cancer cells and assists basophils with
your allergic response.
• Basophils: Produces an allergic response like coughing, sneezing or a runny nose.
• Monocytes: Defend against infection by cleaning up damaged cells