Leukocytes: The Granulocytic and

Monocytic Series
Part I

Chapter 14
The Leukocytes
Learning objectives:
Upon completion of this lecture series the learner should
be able to competently discuss:
The differentiation and maturation of the granulocyte cell
series in detail
The differentiation and maturation of the monocytemacrophage cell series in detail
The function of granulocytes and monocytes
Methods of assessment of granulocytes and monocytes in
a clinical setting (absolute counts, differential counts,
morphology, staining patterns, etc.)

Part I: Granulocytes
Development and proliferation passes through 3 major phases in 3
different sites:
Phases
1. Proliferative phase: for cell number to increase and for
cells to start to differentiate
2. Maturation-storing phase: cells complete their maturation
and replenish the constantly changing reservoir in the BM
storage compartment.
3. Distribution / release to circulation then to final
destination depending on demand which changes; infection
and tissue injury increases demand.
Sites
- Proliferative compartment of BM
- Maturation/storing compartment of BM
- Circulation & the endothelium of the vascular system)

Journey of a leukocyte

Proliferative phase

Occurs in the proliferative compartment (mitotic pool) of BM which

houses cells capable of DNA synthesis & mitosis 4 stages:
Myeloblast: blast denotes enlarged cells undergoing proliferation; represent
the first cell that is morphologically recognizable as a granulocyte (granulation,
N/C ratio, chromatin pattern [clumped vs. dispersed], shape of nucleus, etc.)
Promyelocyte
Myelocyte
Metamyelocyte

Maturation-storing phase:
Within

this

compartment

metamyelocytes

coming

from

proliferative compartment make up to 45% of cells

Mature into band neutrophils (35%) [band describes the shape

of the nucleus in such cells

Further

develops

into

segmented

neutrophils

(20%)

[segmented pertains to shape of nucleus as well].

Note that same phases apply to basophils, eosinophils and

monocyets.

Band vs. segmented neutrophil

Band neutrophil in
peripheral blood film

Segmented neutrophil in
peripheral blood film

Distribution / Release to circulation:

Under the influence of various profiles of interleukins &

concurrent with the expression of homing markers (various
members of the selectin and integrin families like VCAM, ICAM, Lselectin, LFA-1, etc) on vascular endothelium, mature
granulocytes start rolling towards sinusoid walls of BM into
circulation.
Granulocyte movement is also enhanced by cellular changes
decrease in size, N/C decrease, increased flexibility.
In the vascular system cells either stay in circulation or gather at
marginating pools.
Marginating granulocytes adhere to the endothelim of blood
vessels; can be released on demand.
If needed as phagocytes (e.g. tissue injury), granulocytes (mainly
neutrophils) exit circulation to tissues by diapedesis. See next slide

Developmental pathways of granulocytes & monocytes

Maturation of Neutrophils
1. Myeloblast
Earliest neutrophil precursor
Size: 10-18 m
N:C ratio: High(4:1)
Nucleus: Oval or round
Chromatin: delicate, lacy, evenly stained
Nucleoli: 3-5
Cytoplasm: small rim of basophilic cytoplasm
that lacks granules.
Golgi apparatus: unstained area adjacent to
nucleus
CD markers: CD33 and CD38.
Auer rods : which are aggregates of fused
lysosomes, may appear as red, needle-like
crystalline cytoplasmic inclusions.
These inclusions may appear alone or in
groups

Maturation of Neutrophils
2. Promyelocyte
Size: larger than blast (14-20 m)

N:C ratio: High(3:1)

Nucleus: large, Oval or round

Chromatin: Coarser than blasts, lacy, purple-blue

Nucleoli: less prominent or absent

Cytoplasm: pale grayish blue.

Granules: Large blue purple primary granules (azurophilic

granules that contain loads of AMPs = antimicrobial

peptides(myeloperoxidase and chloroacetate esterase.

Maturation of Neutrophils
3. Myelocyte
(The last stage that is capable

of division within BM)

Size: 12 to 18 m
N:C ratio: Decreased(2:1-1:1)

Nucleus: has a more oval appearance than

in previous stages
Chromatin: More condensed than blasts
Nucleoli: Usually absent
Cytoplasm: Light pink, with blue batches
Granules: Secondary granules (specific): small, pink red to
pink violet in color.
The separate cell types: neutrophils, eosinophils, and
basophils become visibly recognizable at this stage.

Maturation of Neutrophils
4. Metamyelocyte
Size: 10 to 18 m
N:C ratio: Decreased
Nucleus: Kidney-bean shaped
Chromatin: coarse, clumped, stains dark purple
Nucleoli: Not visible

Cytoplasm: Neutral pink

Granules: Predominance of
small pinkish red specific granules

Maturation of Neutrophils
5. Band Neutrophil
(also called stab neutrophil,
and may normally present
in peripheral blood).
Size: 10 to 16 m
N:C ratio: Decreased (1:1)
Nucleus: More indented than
metamyelocyte
Horseshoe shape

Chromatin: pyknotic (condensed) at either end

of nucleus, Stain dark purple
Cytoplasm: Pink

Granules: Abundant small pinkish red specific granules

Maturation of Neutrophils
6. Segmented Neutrophil
(Polymorphonuclear)
Size: 10 to 16 m
N:C ratio: Decreased
Nucleus: Segmented, with
two to five lobes connected
by thin filament
Chromatin: condensed, stain deep
purple black
Cytoplasm: Pink
Granules: Mainly small pinkish red
specific granules

Maturation of Eosinophils

Morphological maturation of eosinophils is similar

to that of neutrophils.
Only at the myelocyte stage can the cell be
identified as an immature eosinophil.
Eosinophilic myelocyte
staining granules.

contain

large,

eosin-

Maturation from myelocyte to the metamyelocyte,

band, and segmented eosinophil stage is similar to
neutrophils.

Characteristics of Eosinophils

Size: 12 to 15 m
Nucleus: No more than 2-3
lobes.
Chromatin: Stain dark purple
Cytoplasm: Orange red

Granules: Abundant with large

acidophilic (Eosin-loving) granules.

Maturation of Basophils
Morphological maturation of basophils is similar
to that described for neutrophils.
First recognizable stage is promyelocyte:
basophilic promyelocyte is smaller, with higher
N:C ratio than that of neutrophil or eosinophil
Basophilic promyelocyte contain large purple

black granules.
Maturation from promyelocyte to the

metamyelocyte, band, and segmented basophile

stage is similar to neutrophils.

 Basophilic granules contain

heparin and histamine.

Mast cells have granules that

have an enzyme content
similar to those of the blood
basophil.

Characteristics of Basophils
Size: 10 to 15 m

Nucleus: Segmented
Chromatin: Stain dark

purple
Granules: Many purple

in color, obscuring
the background of
the cytoplasm

Myeloblast: High N/C ratio. Cytoplasm is agranular. Nucleus has a fine

chromatin pattern

Promyelocyte: Cytoplasm contains nucleus with a fine chromatin pattern and

nucleoli. Numerous primary azurophilic granules.

Myelocyte: Cytoplasm is pinkish blue, indicating secondary granule formation.

Nucleus is oval and eccentric with a moderately clumped chromatin pattern.

Metamyelocyte: Cytoplasm contains a few azurophilic primary granules but

mostly pinkish-blue secondary granules. Nucleus indented with dense chromatin
clumping.

Neutrophilic Band: Cytoplasm is pinkish-blue due to secondary granules. Nucleus is

clumped, and has a horseshoe shape that has uniform width throughout the nucleus.

Segmented Neutrophil: Cytoplasm is finely granulated. Nucleus is segmented.

Eosinophilic Myelocyte: Cytoplasm contains numerous red-orange granules and few

dark azurophilic granules. Nucleus is oval and eccentric.

Basophilic Myelocyte: Cytoplasm has smaller black granules that are more intense on
the right of the nucleus. Nucleus is oval with moderately clumped chromatin.

Basophil: Cytoplasm contains numerous large black granules that obscure the nucleus.