HEMATOLOGY
1. Introduction
2. Hematopoiesis
3. Erythrocyte
Formation, Structure, Physiology,
Metabolism, Destruction
4. Overview Of Hemoglobin
Withdraw
blood
• hematocrit - centrifuge blood
to separate components
• erythrocytes are heaviest and
settle first
Centrifuge
• 37% to 52% total volume
Plasma
• 55% of whole blood
• Least dense component
Buffy coat
• Leukocytes and platelets
• <1% of whole blood
Erythrocytes
1 Withdraw 2 Centrifuge the • 45% of whole blood
blood and place blood sample. • Most dense
in tube. component
1. Distribution of
• O2 and nutrients to body cells
• Metabolic wastes to the lungs and kidneys
for elimination
• Hormones from endocrine organs to target
organs
2. Regulation of
• Body temperature by absorbing and
distributing heat
• Normal pH using buffers
• Adequate fluid volume in the circulatory
system
3. Protection against
• Blood loss
• Plasma proteins and platelets initiate clot
formation
• Infection
• Antibodies
• Complement proteins
• WBCs defend against foreign invaders
• 90% water
• Proteins are mostly produced by the liver
• 60% albumin
• 36% globulins
• 4% fibrinogen
PORTLAND
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Sites of production:
• Stem Cells
• Growth Factors
• Microenviroment
Unipotent Lymphocytes
Progenitors
WBC’s
RBC’s
Basophils
Eosinophils
Thrombocytes Erythrocytes
Neutrophils
Monocytes Platelets Erythropoietin
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Platelets Erythrocytes Monocyte
Neutrophils Lymphocyte
Copyright © 2010 Pearson Education, Inc. Figure 17.2
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PowerPoint® Lecture Slides
prepared by Vince Austin,
Bluegrass Technical
and Community College
ERYTHROPOIESIS CHAPTER
• the process of
formation of rbc
• Phases in development
1. Ribosome synthesis
2. Hemoglobin accumulation
3. Ejection of the nucleus and formation of
reticulocytes
• Reticulocytes then become mature
erythrocytes
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• Then from here it develops to reticulocyte and
finally ends up with erythrocyte (mature red
cell).
• The whole process takes about 7 days.
• This process of normoblastic maturation is
characterized by the following progressive
changes:-
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1) The cell size diminishes.
2) Ripening of the cytoplasm. the staining reaction
of the cytoplasm changes, the RNA starts
diminishing) These two things take place
simultaneously.
a) So, color changes to pink from blue due to
reduction of RNA matter such that in
Romanowsky stains,
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there is a change in colour from deep blue to
pink due to progressive formation of acidophilic
staining haemoglobin and the simultaneous
reduction of RNA which is responsible for
basophilic of cytoplasm.
b) Haemoglobin formation.
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3) Ripening (maturing) of the nucleus having the
large reddish, purple, open-network of the
nucleus of a polychromatic normoblast
converted to small deeply staining blue black
structureless nucleus of the orthochromatic
normoblast before it is eventually lost. i.e. the
nucleus of the pronormoblast is larger than
that of the orthochromatic normoblast.
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• The former (pronormoblast) has structures and
stains reddish purple, while the later
(orthochromatic) stains blue-black and is
structure less.
• Mitotic division of the developing cells in this
series occurs up to the stage of the
polychromatic normoblast and therefore the
orthochromatic normoblast is not considered
capable of mitotic division.
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Regulation of Erythropoiesis
DECREASE IN ARTERIAL
OXYGEN
DECREASE IN TISSUE
OXYGEN
RELEASE OF ERYTHROPOIETIN
BONE MARROW
ERYTHROPOIESIS
CORRECTION OF HYPOXEMIA
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Hormonal Control of Erythropoiesis
• Erythropoietin (EPO)
• Direct stimulus for erythropoiesis
• Released by the kidneys in response to
hypoxia
• Causes of hypoxia
• Hemorrhage or increased RBC destruction
reduces RBC numbers
• Insufficient hemoglobin (e.g., iron deficiency)
• Reduced availability of O2 (e.g., high altitudes)
• Effects of EPO
• More rapid maturation of committed bone
marrow cells
• Increased circulating reticulocyte count in 1–
2 days
• Testosterone also enhances EPO production,
resulting in higher RBC counts in males
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• This hormone acts on erythroid responsive
cell.
• The cytoplasm is more and stains deep
blue.
• The nucleus is round and occupies most of
the cell and has one or two nucleoli.
• The cell measures 12-20 micrometre in
diameter.
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Proerythroblast
(Pronormoblast/ Rubriblast)
• No hemoglobin
• Nucleus 12 um
• Contain nucleoli
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• The chromatic strands are more thicker and
more deeply staining living a coarser
appearance due to RNA.
• At this stage the nucleus have disappeared.
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Basophil erythroblast (Prerubricyte)
• Early normoblast
• Nucleoli disappear
• Show mitosis
• Increase in RNA
• Hemoglobin starts
appearing – Little Hb
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3. Polychromatic normoblast
(intermediate)
• They vary from 8/14 micrometer in diameter.
• The nucleus is smaller and occupies a smaller
part of the cell.
• It is reddish purple in colour.
• The cytoplasm is large and begins to acquire
haemoglobin, thus taking acidophillic – pink or
pale pink colour.
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Polychromatophil erythroblast (Rubricyte)
• Late normoblast
• Nucleus smaller
• Coarse Chromatin
• Hemoglobin increase
• Eosinophil Stain
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• The cytoplasm is typically acidophillic (pink)
because haemoglobinization has started
taking place which gives it acidophillic
staining.
• The nucleus is lost at this stage.
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Orthochromatic Erythroblast
(Metarubricyte)
• Normoblast
• Nucleus smaller
• Pyknosis
• Nuclear extrusion
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5. Reticulocytes
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• The Hb content is almost the same as that of
mature RBC.
• From the stage of retic to mature RBC takes
about 1-2 days.
• The nucleus is injected out at the spleen and
the RNA and any other inclusions are removed
and then the cell is released in to the
circulation in the blood stream.
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• As the cell matures it becomes smaller in size
and the nucleus diminishes and eventually lost
i.e. the chromatin becomes less dense and the
nucleus diminishes and eventually lost.
• Nucleoli disappear before the nucleus is lost.
• At this stage the cytoplasm losses the protein
synthesis property and acquires haemoglobin.
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Reticulocyte
• Reticulum
• Remnant of ER & GA
• Synthesize Hb
• Few Mitochondria
• 1 % of Red Cells
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ERYTHROCYTE
Basophilic Orthochromic
Polychromatophilic
Pronormoblast Normoblast Normoblast Normoblast
1 Stimulus:
Hypoxia (low blood
5 O2- carrying O2- carrying ability)
ability of blood due to
increases. • Decreased
RBC count
• Decreased amount
of hemoglobin
• Decreased
availability of O2
4 Enhanced
erythropoiesis
increases RBC
2 Kidney (and liver to
a smaller extent)
count.
releases
3 Erythropoietin erythropoietin.
stimulates red
bone marrow.
1 Stimulus:
Hypoxia (low blood
O2- carrying ability)
due to
• Decreased
RBC count
• Decreased amount
of hemoglobin
• Decreased
availability of O2
1 Stimulus:
Hypoxia (low blood
O2- carrying ability)
due to
• Decreased
RBC count
• Decreased amount
of hemoglobin
• Decreased
availability of O2
1 Stimulus:
Hypoxia (low blood
O2- carrying ability)
due to
• Decreased
RBC count
• Decreased amount
of hemoglobin
• Decreased
availability of O2
1 Stimulus:
Hypoxia (low blood
O2- carrying ability)
due to
• Decreased
RBC count
• Decreased amount
of hemoglobin
• Decreased
availability of O2
4 Enhanced
erythropoiesis
increases RBC
2 Kidney (and liver to
a smaller extent)
count.
releases
3 Erythropoietin erythropoietin.
stimulates red
bone marrow.
1 Stimulus:
Hypoxia (low blood
5 O2- carrying O2- carrying ability)
ability of blood due to
increases. • Decreased
RBC count
• Decreased amount
of hemoglobin
• Decreased
availability of O2
4 Enhanced
erythropoiesis
increases RBC
2 Kidney (and liver to
a smaller extent)
count.
releases
3 Erythropoietin erythropoietin.
stimulates red
bone marrow.
4 New erythrocytes
enter bloodstream;
5 Aged and damaged function about 120 days.
red blood cells are
engulfed by macrophages
of liver, spleen, and bone
marrow; the hemoglobin Hemoglobin
is broken down.
Heme Globin
Circulation
Food nutrients,
including amino acids, 6 Raw materials are
Fe, B12, and folic acid, made available in blood
are absorbed from for erythrocyte synthesis.
intestine and enter
blood.
4 New erythrocytes
enter bloodstream;
function about 120 days.
Circulation
Circulation
Food nutrients,
including amino acids, 6 Raw materials are
Fe, B12, and folic acid, made available in blood
are absorbed from
for erythrocyte synthesis.
intestine and enter
blood.
4 New erythrocytes
enter bloodstream;
5 Aged and damaged function about 120 days.
red blood cells are
engulfed by macrophages
of liver, spleen, and bone
marrow; the hemoglobin Hemoglobin
is broken down.
Heme Globin
Circulation
Food nutrients,
including amino acids, 6 Raw materials are
Fe, B12, and folic acid, made available in blood
are absorbed from for erythrocyte synthesis.
intestine and enter
blood.
Factors:
• Growth inducers
• Differentiation inducers.
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ERYTHROPOIESIS
PHSC
Bone marrow
CFU-E
Polychromatophil erythroblast
Orthochromatophil erythroblast
By
Diapedesis
(squeezing through the pores of the capillary
membrane).
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RBCs
Form, Function, and Formation
7.5 µm
Top view
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Erythrocytes
The erythrocyte is not an inert body but a self supporting cell capable
of existing in the intravascular circulation for up to 120 days
Performs extremely complex and important homeostatic maneuvers
A simple structure in that it posses only a simple
membrane, internal stroma, and 90% hemoglobin
Biconcave shape allows maximal surface area and
greatest flexibility
The membrane is bilayered with a lipid to protein ratio
of 1:1.6 the majority being lipoproteins
Semipermeable to water
Stroma is composed of lipids and proteins in the form of
fibrous proteins
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THE ERYTHROCYTE
Metabolism
Metabolism is basically:
89% Embdden-Meyerhoff
11% Pentose phosphate pathway
The process of hemolysis is favored when the pathways
fail to provide sufficient reduced glutathione or sufficient
energy-providing substances (NADH, NADPH or ATP)
About 20 chemicals are involved in the pathways
• Na + - K + pump
• Iron in Fe ++ form
• Functions:
• Rbc’s critical role is in the transport of oxygen and
carbon dioxide and is dependent on hemoglobin
• CARBONIC ANHYDRASE– enzyme in the rbcs that
catalyzes the reaction that joins carbon dioxide and
water to form carbonic acid
• CARBONIC ACID– dissociates and generates
bicarbonate ions, which diffuses out of the rbcs and
serves to transport carbon dioxide in the blood plasma
• Hemoglobin structure
• Protein globin: two alpha and two beta chains
• Heme pigment bonded to each globin chain
• Iron atom in each heme can bind to one O2
molecule
• Each Hb molecule can transport four O2
Heme
group
Globin chains
(a) Hemoglobin consists of globin (two (b) Iron-containing heme pigment.
alpha and two beta polypeptide
chains) and four heme groups.
5’ 3’
5’ 3’
1. Insufficient erythrocytes
• Hemorrhagic anemia: acute or chronic loss
of blood
• Hemolytic anemia: RBCs rupture
prematurely
• Aplastic anemia: destruction or inhibition of
red bone marrow
• Pernicious anemia
• Deficiency of vitamin B12
• Lack of intrinsic factor needed for absorption
of B12
• Treated by intramuscular injection of B12 or
application of Nascobal
3. Abnormal hemoglobin
• Thalassemias
• Absent or faulty globin chain
• RBCs are thin, delicate, and deficient in
hemoglobin
• Sickle-cell anemia
• Defective gene codes for abnormal
hemoglobin (HbS)
• Causes RBCs to become sickle shaped in
low-oxygen situations