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General Cell Maturation

Characteristics
IMMATURE CELLS

MATURE CELLS

Cell is large
Nucleoli are present

Cell becomes smaller


Absent Nucleoli

Chromatin - fine and


delicate

Coarse and clumped

Nucleus - round

Round, lobulated or
segmented

Cytoplasm dark blue


(rich in RNA)

Light blue (less RNA)


cytoplasm

High N:C
Low N:C ratio
(nucleus:cytoplasm) ratio

Erythrocyte Maturation
Erythropoiesis red cell production and

maturation
CD34 hematopoietic stem cell marker
Erythropoiesis is regulated by
erythropoietin, which is produced by
the kidneys
Strongest stimulus for erythropoietin
production: tissue hypoxia

Erythrocyte Maturation

Erythrocyte Maturation
Maturation Stages:
Six morphological stages of erythrocyte maturation

may be identified with Wright stain


Normal maturation is dependent on intake of
proper nutrients and vitamins such as folate,
vitamin B12 and iron
Nomenclature:
There are three nomenclatures used to describe
the six stages
Rubri (proposed by the ASCP)
Erythroblast (proposed by Paul Ehrlich)
Normoblast (normal precursor)

Erythrocyte Maturation

Erythrocyte Maturation
General Guidelines:
The ASCP published the following general guidelines

for identification of erythroid precursors:


Progressive decrease in size and the degree of
cytoplasmic basophilia (blue color) as the cell
matures
Nuclei are round or oval in the blast stage become
round thereafter
Gradual increase in coarseness and condensation of
the chromatin, ranging from fine in the early stages
to pyknotic in the stage just before nuclear extrusion

Rubriblast
Pronormoblast
) m,
This is the earliest(erythrocyte,
cell size up to 20
The nuclear:cytoplasmic ratio is 8:1
1-3 nucleoli, nucleus has dark areas of DNA
Chromatin is fine and uniform and stains intensely
Deep blue cytoplasm with no granules

Prorubricyte (Basophilic
normoblast)

Size up to 16 m with an N:C ratio of 4:1


Centrally located nucleus with 0-1 nucleoli
Chromatin coarsening
Cytoplasm is less blue but intensely basophilic

RNA

Rubricyte (Polychromatophilic
normoblast)

Size up to 12 m with an N:C ratio of 4:1


Eccentric nucleus with no nucleoli
Chromatin shows significant clumping
Hemoglobin synthesis starts in this stage
The RNA and hemoglobin give the cytoplasm a

reddish-blue color called polychromasia or


polychromatophilia

Metarubricyte (Orthochromic
normoblast)

This is the last nucleated erythrocyte stage


Size up to 10 m with an N:C ratio of 0.5:1
Eccentrtic nucleus with small, fully condensed

(pyknotic) nucleus; no nucleoli


Pale blue to salmon cytoplasm
Hemoglobin synthesis decreases

Reticulocyte (diffusely basophilic


erythrocyte)
Size up to 10 m
Contains no nucleus but has mitochondrial and ribosomes
Last stage to synthesize hemoglobin
Last stage in bone marrow before release to the blood
Supravital stain is used for identification
One of the best indicators of bone marrow function
Hemoglobin continues to be produced for approx 24 hours

after exiting the bone marrow

Mature Erythrocyte

The mature erythrocyte is approximately 6-8 m in

size
It is a biconcave disc and hence referred to as a
discocyte
In a Wright stain, a central pale area is revealed which
fades gradually into the reddish pink cytoplasm

Granulocyte
Maturation

MYELOBLAST

Earliest recognizable granulocyte precursor


14-20 m; NC ratio 7:1-4:1
Round/oval nucleus with fine reddish-purple

staining chromatin
2-5 nucleoli; dark blue cytoplasm
No cytoplasmic granules
1% of the nucleated cells in the bone marrow

PROMYELOCYTE

15-21 m; NC ratio 3:1


Round/oval nucleus with slightly coarsening

chromatin
1-3 nucleoli; dark blue cytoplasm
Cytoplasm has large, nonspecific/primary
granules containing myeloperoxidase
2-5% of the nucleated cells in the bone marrow

MYELOCYTE

First stage where the different granulocyte types can

be differentiated
12-18 m; NC ratio 2:1; round nucleus with coarse chromatin
Light blue to light pink cytoplasm; prominent golgi apparatus
Specific/secondary granules that contain hydrolytic enzymes
(e.g. alkaline phosphatase and lysozyme) are present
Nonspecifc granules are present and may still stain
Last stage capable of cell division

METAMYELOCYTE

10-18 m; NC ratio 1.5:1


Nucleus is indented in a kidney bean shape and has

coarse, clumped chromatin


Nuclear indent less than half the width of a
hypothetical round nucleus
Cytoplasm is pink filled with specific/secondary granules
Primary granules are present but do not stain

BAND (STAB)

10-15 m; NC ratio 1:2


Nucleus is C or S-shaped with coarse, clumped chromatin
Nuclear indent is greater than half the width of a hypothetical round

nucleus
Cytoplasm is pink and filled with specific/secondary granules
Primary granules are present but do not stain
Stored in the bone marrow and released when there is increased
demand
Earliest stage that can be seen in the periperal blood of a
healthy person

MATURE GRANULOCYTES

NEUTROPHIL EOSINOPHIL
10-15 m; NC ratio
1:3

12-16 m

BASOPHIL
10-15 m

Nucleus has coarsed,


clumped chromatin
with 3-5 lobes
connected by thin
filaments

Nucleus is usually
bilobed

Nucleus is
unsegmented or
bilobed

Cytoplasm contains
fine lilac secondary
granules

Cytoplasm contains
large, bright redorange secondary

Cytoplasm contains dark


violet to purple blue
secondary granules;