QUIZ 1A HEMA LEC b.

Slightly alkaline
1. What is the name of the iron-containing protein c. Composed of mostly WBCs and platelets
that give red blood cells their color? d. Cooler than body temperature
a. Hemoglobin 11. Which of the ff. statements are true?
b. Myoglobin a. All statements are true
c. Hemocyanin b. Blood consists of plasma and formed elements
d. Pyrite c. Plasma is approximately 92%
2. A decrease in the oxygen-carrying capacity of the d. Plasma is straw colored clear liquid containing
blood for any reason, is a condition known as cellular elements and solutes
a. Leucocytosis 12. Function of blood includes
b. Polycthemia a. Maintains body temperature
c. Thrombocytopenia b. Defend against infection
d. Anemia c. All the options are correct
3. Which of the following WBC is capable of d. Transport oxygen
phagocytosis? 13. What is the normal pH of the blood?
a. Lymphocyte a. 7.30-7.40
b. Neutrophil b. 7.20-7.30
c. Basophil c. 7.45-7.55
d. Eosinophil d. 7.35-7.45
4. Which if the ff. statements about erythrocytes is 14. The major components of plasma is
correct? a. Gases
a. They lack a nucleus b. Proteins
b. They are produced in the spleen c. Nutrients
c. They clot blood d. Water
d. They fight infection 15. The most abundant formed elements in the blood
5. Which of the following is the function of WBC? a. Globulins
a. Transport oxygen b. Leukocytes
b. Defend against infection c. Erythrocytes
c. Maintain homeostasis d. Albumins
d. Produce haemoglobin 16. Erythrocytes
6. Which of the following leukocytes have the ability a. Contain large quantities of haemoglobin
to produce antibodies? b. Are biconvex disks
a. B lymphocytes c. Divide frequently
b. Erythrocytes d. Have several nuclei in each cell
c. Monocytes 17. The WBC called granulocytes are
d. Neutrophils a. Monocytes, leukocytes, and neutrophils
7. Which of the ff leukocytes respond when a person b. Lymphocytes and monocytes
is allergic to something? c. Neutrophils, basophils, and eosinophils
a. Basophils d. Erythrocytes, thrombocytes, and platelets
b. Neutrophils 18. What is haematocrit?
c. Eosinophils a. a medicine that helps the bleeding stop
d. Lymphocytes b. a sometime fata blood disease
8. What is the fluid portion of the unclotted blood c. the proportion of RBC compared with total blood
called? volume
a. Neither plasma nor serum d. a blood pressure measuring device
b. Both plasma and serum 19. Mean cell haemoglobin concentration is increased in
c. Serum which of the ff. conditions?
d. Plasma a. Megaloblastic anemia
9. What are the little cellular fragments called that b. Spherocytosis
are responsible for clotting blood? c. All the options are correct
a. Thrombocytes d. Macrocytic anemia
b. Erythrocytes
c. Leukocytes
d. Haemoglobin 20. An indicator of the average weight of haemoglobin
in individual cell
10. Blood is a. MCHC
a. Acidic b. MCH
c. RDW
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 d. MCV HEMA LEC QUIZ 2 LEC (25/25)
21. The parameters required in calculating the RBC 1. All of the ff. do not occur in the medullary phase,
indices include the ff. except EXCEPT
a. Haematocrit o Recognizable clusters of developing
b. Haemoglobin erythroblasts, granulocytes and monocytes begin
c. RBC count to form
d. WBC count o Yolk sac begins to disappear
22. Mean cell volume is decreased in which of the ff. o Myeloid:erythroid ratio reaches 3:1
conditions? o Production of gower 1 and 2 hemoglobin
a. Iron deficiency anemia 2. Which of the ff. statements might NOT be true?
b. Haemolytic anemia o In adults, the thymus is virtually atrophied and is
c. Liver disease therefore no longer functional
d. Megaloblastic anemia o Not all lymphocytes are leukocytes
23. The normal reference for WBC - Correct answer: All lymphocytes are
a. 4800-10,800 cells leukocytes but all leukocytes are not
b. 3000-7000 cells lymphocytes.
c. 150,000-400,000 cells o Pluripotent hematopoietic stem cells are
d. 4-6 million cells produced in the bone marrow
24. If there is an increased RBC Count leading to o Thymocytes are educated in the thymus
hyperviscosity of blood, the condition is known as: 3. Which organ do B lymphocytes mature?
not sures o Spleen
a. None of the options are correct o Bone marrow
b. Leukimia o Thymus
c. Anemia o Lymph nodes
d. Polycythemia 4. Which of the ff. cells surround the cavity of the yolk
25. Volume of blood in averaged size adult male is sac?
a. 6 to 7L o Erythroblasts
b. 5 to 6L o Angioblasts
c. 3 to 4L o Lymphoblasts
d. 4 to 5L o All of the cells are correct
5. Cellularity is described in adults as hypoplastic when
the
o The marrow has <30% hematopoietic cells
o The marrow has >70% hematopoietic cells
o The marrow has >30% hematopoietic cells
o The marrow has <70% hematopoietic cells
6. The paracortical area of the lymph node is occupied
by the _____ cells
o T
o All of the cells are correct
o B
o Natural killer
7. Which of the ff. is not true of medullary phase of
hematopoiesis?
o Begins in the bone marrow cavity
o Measurable levels of erythropoietin can be
detected
o Hematopoietic activity apparent
o Production of megakaryocytes begins in this
phase
8. Myeloid:erythroid ration in normal adult is
o 6:1
o 5:2
o 3:1
o 5:1
9. Which of the ff. cells is a product of the common
lymphoid progenitor?
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 o Granulocyte o All of the items are correct
o Megakaryocyte o Erythropoetin
o T lymphocyte o Hemoglobin F and A
o Erythrocyte 19. The lymph is the fluid portion of blood that escapes
10. Which organ is the site of sequestration of into the connective tissue and is characterized by
membrane damaged RBCs and removes them from o Both statements are correct
the circulation? o A low protein concentration
o Spleen o None of the options are correct
o Thymus o The absence of RBCs
o Bone marrow 20. During the second trimester of fetal development,
o Liver the primary site of blood cell production is the
11. In severe hemolytic anemias, the liver ____ the o Bone marrow
conjugation of bilirubin and ____ the storage of o Liver
iron o lymph node
o Increases, decreases o spleen
o Decreases, increases 21. culling and pitting are functions of
o Decreases, decreases o Lymph node
o Increases, increases o Bone marrow
12. Progenitor cell which can give rise to mast cells o Spleen
o Common lymphoid progenitor o Liver
o CFU-GEMM 22. Which of the following forms an extracellular matrix
o Common myeloid progenitor promoting cell adhesion and regulating
o Pluripotent stem cells hematopoietic stem cells?
13. Theory stating that all blood cells come from one o Stromal cells
origin stem cell o Mesenchymal cells
o Monophyletic o Reticular cells
o Instructive o Epithelial cells
o Stochastic 23. The site for primitive hematopoiesis?
o Polyphyletic o Bone marrow
14. Which organ is the site of sequestration of o Spleen
platelets? o Yolk sac
o Spleen o Liver
o Bone marrow 24. The process of formation and development of blood
o Liver cells is termed
o Thymus o Hemocytometry
15. As one ages, the size of the thymus o Hematopoiesis
o Stays the same o Hematorrhea
o Decreases o Hematemesis
o Depends on the rate of the development of 25. Active red marrow hematopoiesis occurs in the
infection following, EXCEPT
o Increases o Skull
16. Which of the following cells are found in the liver o Iliac crest
acting as macrophages? o Sternum
o Langerhan cells o Kidney
o All cells are correct
o microglial cells
o Kupffer cells
17. Which of the following are functions of the
hepatocytes in the liver?
o Coagulations factors synthesis
o All statements are correct
o Carbohydrate and lipid metabolism
o Protein synthesis and degradation

18. Which of the following are detected during the

hepatic phase of hematopoiesis?
o G-CSF
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QUIZ 3 HEMA LEC b. None
1. Most immature erythrocyte precursor? c. Parachromatin
a. Myeloblast d. Heterochromatin
b. Metamyelocyte 11. All are characteristic of the myeloblast except that
c. Pronormoblast it has
d. Promyelocyte a. Nucleoli
2. As the erythrocyte mature, the nucleolus b. Low N:C ratio
cytoplasmic ratio c. Large nucleus with fine chromatin
a. increases d. Dark blue, scantly cytoplasm without granules
b. cannot be determined 12. Which of the ff. morphologic changes occurs during
c. decreases normal blood cell maturation?
d. stays the same a. Condensation of nuclear chromatin
3. What is the youngest cell in the maturation series b. Appearance of nucleoli
of the neutrophils? c. Increase in cell cytoplasm
a. Promyelocyte d. Development of cytoplasm basophilia
b. Metamyelocytes 13. The metamycelocyte matures to this type of cells
c. Myeloblast a. Monocyte
d. Myelocyte b. Band neutrophil
4. Which of these is true regarding the maturation of c. Myelocyte
RBC precursors? d. Basophil
a. The proportion of nucleus increases; the 14. The cytoplasm of the blast cell tends to strain much
cytoplasmic proportion decreases darker blue because of
b. Cytoplasmic color change from pink to gray to a. Increased amount of DNA
blue b. Increased amount of specific granules
c. Nucleoli becomes more prominent c. Increased amount of RNA
d. The nuclear chromatin becomes coarser, d. Increased amount of azurophilic granules
clumped and condensed. 15. Which of the ff. involves in asynchronistic
5. The basis for determining cell’s maturity maturation of cells?
a. Neither the nucleus nor the chromatin a. Cytoplasm
b. Both the nucleus and chromatin b. All items are correct
c. Nucleus c. Nucleus
d. Chromatin d. Size
6. Euchromatin is described as 16. Classification test
a. Very delicate, fine, and linear Cell is large – immature cells
b. Increasingly coarse and clumped Nucleoli present – immature cells
c. Decreasingly coarse and clumped Low N:C ration – mature cells
d. Non-staining areas in the nucleus of old cells Chromatin coarse and clumped – mature cells
7. All of these are changes observed in the cytoplasm Cytoplasm rich in RNA – immature cells
of hematopoetic stem cells except
a. Increase in proportion (vs nucleus)
b. Increase in basophlia
c. Change in color
d. Possible appearance of granules
8. Which type of leukocyte can myeloid progenitor
cells not produce?
a. B cells
b. Eosinophils
c. Platelets
d. Monocytes
9. Characteristic of the myeloblast
a. Has dark blue, scanty cytoplasm
b. All the statements are correct
c. Has large nucleus with fine chromatin network
d. Has nucleoli
10. The non staining areas in the nucleus of old cells
are called
a. Euchromatin
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MT-HEMA 1.1 (Hematology) LEC  Blood volume is 5L to 6L (1.5 gal) in an
average sized adult male and 4L to 5L (1.2
Historical Developments and General Characteristics gal) in an average sized adult female
of Blood  Newborn’s blood volume in 200-250mL of the
total blood volume
Hematology  Normovolemia – normal blood volume
 Concerned with the diseases of blood and blood-  Hypervolemia – increased blood volume
forming tissues o Excessive fluid intake
o Blood transfusion
Blood o IV injection of fluids
 Vital, life-sustaining fluid circulating in a closed system  Hypovolemia – decreased blood volume
of blood vessel and the heart o Loss of blood (bleeding or hemorrhage)
 The systemic circulation provides the functional blood o Loss of erythrocytes (hemolytic anemias)
supply to all body tissue. It carries oxygen and o Loss of plasma (severe burns)
nutrients to the cells and picks up carbon dioxide and o Loss of body fluids (diarrhea, LBM,
waste products. excessive sweating)
 It carries oxygenated blood from the left ventricle  Functions
through the arteries to the capillaries in the tissues of - Transportation
the body  Respiration
 Characteristics  Nutrient carrier from GIT
- Color  Transportation of hormones from endocrine
 Arterial – bright red or bright scarlet red glands
color (high oxygen content)  Transports metabolic wastes
 Venous – very dark red (low oxygen content) - Regulation
- Viscosity  Regulates pH
 Whole blood – 4.5 to 5.5 times as viscous as  Adjusts and maintains body temperature
water, more resistant to flow than water  Maintains water content of cells
- Reaction - Protection
 Alkaline pH 7.35-7.45, average 7.4  WBC protects against disease by phagocytosis
 pH is controlled by the buffer system of the  Reservoir for substances like water,
blood electrolyte, etc.
- Temperature - 38°C (100.4°F)  Performs haemostasis
- Osmolality  Composition
 Measure of how much one substance has - Whole blood includes
dissolved in another substance. The greater erythrocytes, leukocytes,
the concentration of the substance dissolved, platelets, and plasma.. when a
the higher the osmolality specimen is centrifuges,
 Very salty water has higher osmolality than leukocytes and platelets make up
water with just a tint of salt. the buffy coat (small layer of
 A normal result is typically 275 to 295 cells lying between the packed
milliosmoles per kilogram. RBCs and the plasma)
 Increased or decreased osmolality calls for - Plasma is the liquid, cell-free portion of unclotted
evaluation of the patient’s fluid and blood that has been treated with anticoagulants.
electrolyte balance. Serum is the fluid that remains after coagulation,
 The normal range for serum osmolality is therefore devoid of clotting factors such as
280-300 mOsm/kg. fibrinogen.
 Above normal values may indicate conditions
such as dehydration, hyperglycemia, diabetes
insipidus, hypernatremia, uremia and renal
tubular necrosis
- Specific gravity – ratio of the density of a
substance to that of a standard substance (water)
 Whole blood – 1.052-1.061
 Plasma – 1.022-1.026
- Volume/amount
 RBCs and plasma, regulated by kidney
 20% of the extracellular fluid amounting to
8% of the total body mass

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- Blood Plasma
 When the formed elements are removed from
the blood, straw-colored liquid called plasma
is left. Plasma 91.5% water and 8.5% solutes,
much of which (7%) are proteins.
 Some of the proteins in plasma are also found
elsewhere in the body but those confined to
the blood are called plasma proteins.
 These proteins play a role in maintaining
proper blood osmotic pressure which is
important in total body fluid balance.
 Most plasma proteins are synthesized by the
liver, including the albumins (54% of plasma
proteins), globulins (38%) and fibrinogen
(7%).
- Formed elements
 They are so named because they are enclosed
in a plasma membrane and have a definite
structure and shape.
 All formed elements are cells, broadly
classified as RBCs, WBCs, except for platelets,
which are tiny fragments of bone marrow
cells.
Red blood cells (erythrocytes)
 Anucleate, biconcave, discoid cells filled with a reddish
protein, hemoglobin, which transports oxygen and
carbon dioxide
 Appear pink to red and measure 6-8µm in diameter
 They have a zone of pallor that occupies one-third of
their center
 RBCs counted in measured volumes can detect anemia
or polycythemia
 Anemia – loss of oxygen-carrying capacity and is often
reflected in a reduced RBC count or decreased RBC
hemoglobin concentration.
 Polycythemia – increased RBC count reflecting
increased circulating RBC mass, a condition that leads
to hyperviscosity.
 RBC indices
- For the computation of RBC indices, three
numerical results—RBC count, Hgb and Hct
(packed cell volume) may be used.
- Part of the complete blood count (CBC) test. They
are used to help diagnose the cause of anemia, a
condition in which there are too few RBCs.
- MCV (Mean corpuscular volume)
 Reference range (SI/Conventional units) is
80-95 femtoliters (fL), newborn 96-180 fL
 An indicator of the average/mean volume of
RBCs
 Reflects RBC diameter on a Wright-stained
blood film
 Calculate using the hematocrit (Hct) and RBC
count
��� (%) 10
MCV (fL) =��� ������10 ( 2)
L
 Low MCV indicates microcytic (small average
RBC size) normal MCV indicates normocytic
(normal average RBC size), and high MCV
indicates macrocytic (large average RBC size)
 Increased MCV can be seen in megaloblastic
anemia, hemolytic anemia with
reticulocytosis, liver disease and normal
newborn.
 Decreased MCV can be seen in iron
deficiency anemia, thalassemia, sideroblastic
anemia and lead poisoning.
- MCH (Mean Corpusular Hemoglobin)
 Reference range (SI/conventional units) is
26-34 picograms (pg)
 An indicator of the average weight of
hemoglobin in individual RBCs
(hemoglobin amount per RBC)
 Expresses the mass of hemoglobin and
parallels the MCHC
 Calculate using the hemoglobin (Hgb) and
RBC count
�� (�/��)� 10
MCH (pg) = �102
��� ����� ( L )
 Increased MCH can be seen in macrocytic
anemia
 Decreased MCH can be seen in microcytic,
hypochromic anemia
- MCHC (Mean Corpuscular Hemoglobin
Concentration)
 Reference range (SI/conventional units) is
32-37 g/dL (SI units) 320-370 g/dL
 A measure of the average concentration of
Hgb in g/dL
 Reflects RBC staining intensity and amount of
central pallor
 Calculate using Hgb and Hct values
��� (�/��) 100
MCHC (g/dL) =
���
 32-37 g/dL MCHC indicates normochromic
RBCs
 Lesser then 32 g/dL MCHC indicates
hypochromic RBC, which is seen in iron
deficiency anemia
 Greater than 37 g/dL MCHC indicates possible
error in RBC or Hgb measurement, or the
presence of spherocytes
 White blood cells
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 Other RBC parameters
- RDW (RBC distribution width)
 A measurement of the range in the volume
and size of the RBCs
 A normal range for RDW is 12.2-16.1& in
adult females and 11.8-14.5 in adult males. If
you score outside this range, you could have a
nutrient deficiency, infection, or other
disorder. However, even at normal RDW
levels, you may still have a medical condition
 Determined from RBC histogram, a graphic
representation of particle size distribution
 High RDW: it could be an indication of
nutrient deficiency of iron, folate or vitamin
B12. These results could also indicate
macrocytic anemia when your body doesn’t
produce enough red blood cells.
 A person may have high RDW and normal
MCV. This suggests a deficiency of iron, B12
or folate. It may also indicate chronic liver
disease. High RDW and low MCV.
 A low RDW means your RBCs are all about
the same size
- Hematocrit
 Reference range for males (conventional
units is 41-53 L/L) (SI units is 0.41-0.53 L/L),
for females (conventional units is 36-46%) (SI
units 0.36-0.46 L/L). Reference range is age
and sex dependent.
 The percentage of RBCs in a given volume
of whole blood.
 The buffy coat layer of leukocytes and
platelets, not included in the measurement,
can be seen between plasma (upper) and RBC
(lower) layers
 Calculate by many automated cell counters
using the MCV and RBC count
��� (��) ��� ����� (�
�102
)
Hct (%) = L disorder
10
- Hemoglobin
 A protein in RBCs that carries oxygen.
 Reference range for males (conventional
units is 13.5-17.5 g/dL) (SI unita 135-175
g/L); for females (conventional units – 12.0-
16.0 g/dL) (SI units – 120-160 g/dL)
 Reference range for hemoglobin is age and
sex dependent.
 High levels could be indicative of the rare
blood diseases, polycythemia. It causes the
body to make too many RBCs, causing the
blood to be thicker than usual.
 This can lead to clots, heart attacks, and
strokes. It is a serious, lifelong condition that
can be fatal if it’s not treated
 Parameters
- All these parameters—RBC count, Hgb, Hct,
indices and RBC morphology—are employed to
detect, diagnose, assess the severity and monitor
the treatment of anemia, polycythemia and the
numerous systemic conditions that affect RBCs.
- Automated hematology profiling instruments are
used in nearly all laboratories to generate these
data, although visual examination of the Wright-
stained blood film is still essential to verify
abnormal results
 Reticulocytes
- Polychromatic (polychromatophilic) erythrocytes,
newly released from the RBC production site: the
bone marrow
- Indicate the ability of the bone marrow to
increase RBC production in anemia due to
blood loss or excessive RBC destruction
White Blood Cells (Leukocytes)
 Leukocytes protect their host from infection and
injury.
 They are transported in the blood from their source,
usually bone marrow or body cavity destination.
 WBCs are so named because they are nearly colorless
in an unstained cell suspension
 Leukopenia – decreased WBC count (fewer than
4500µL)
 Leukocytosis – increased WBC count (more than
11,500µL)
Platelets (Thrombocytes)
 Thrombocytes are true blood cells that maintain blood
vessel integrity by initiating vessel wall repairs.
 They are only 2-4µm in diameter, round or oval,
anucleate “cell fragments”
 They appear insignificant (small size), but they are
essential to life and are extensively studied for their
complex physiology.
 Thrombocytosis – elevated platelet count
 Thrombocytopenia – low platelet count
 Thrombocythemia – life-threatening hematologic
 PLT (platelets)
- Reference range (SI units – 150-450x109/L)
(conventional units – 150,000-450,000/µL)
 MPV (Mean Platelet Volume)
- Reference range (SI/conventional units – 6.8-
10.2 fL)
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 Analogous to MCV for erythrocytes Relative and Absolute Blood Cell Counts
 Relative count – amount of a cell type in relation to
other blood components
- Relative polycythemia – an apparent rise of the
erythrocyte level in the blood, however the
underlying cause is reduced plasma volume
 Absolute count – actual number of each cell type
without respect to other blood components
- Absolute polycythemia – true increase in red cell
mass from any cause
 Complete Blood Count – CBC is performed on
automated hematology profiling instruments and
includes the RBC, WBC and platelet measurements
- RBC parameters
 RBC count
 Hgb
 Hct
 MCV
 MCH
 MCHC
 RDW
 Retic
- Platelet parameters
 PLT count
 MPV
- WBC parameters
 WBC count
 Neutrophil count: % and absolute
 Lymphocyte count; % and absolute
 Eosinophil and Basophil counts: % and
absolute

Basic Homeostasis Basic Hematology Terminology

 Homeostasis – body’s tendency to move toward  a- – without
physiological stability. In vitro testing of blood and  -blast – youngest/nucleated
other body fluids must replicate exact environmental  -chromic – colored
body conditions. These conditions should include the  -cyte – cell
following:  dys- – abnormal
- Osmotic concentration – the body/cellular water  -emia – in the blood
concentration, composed of 0.85% sodium  ferro- – iron
chloride. This normal osmotic concentration is  hyper- – increased
termed isotonic  hypo- – decreased
- Hypotonic solution – greater amount of water in  iso- – equal
relationship to lesser amount of solutes—water  macro- – large
enters the cell; the cell swells and may lyse
 mega- – very large, huge
- Hypertonic solution – lesser amount of water in
 micro- – small
relationship to greater amount of solutes – water
leaves the cell; cell may crenate  myelo- – marrow
 normo- – normal
 -oid – like
 -osis – increased
 pan- – all
 -penia – decreased
 -plasia – formation
 -poiesis – cell production
 poly- – many
 pro- – before
 thrombo- – clot

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Formed Elements and Sizes
Hematopoiesis
Hematopoiesis
 Hematopoiesis is a continuous regulated process of
blood cell production that includes cell renewal,
proliferation, differentiation and maturation
 These processes result in the formation, development
and specialization of all the functional blood cells that
are released from the bone marrow to the circulation
 In healthy adults, hematopoiesis is restricted primarily
to the bone marrow
 During fetal development, the restricted sequential
distribution of cells initiates in the yolk sac and then
progresses in the aorta-gonad mesonephros (AGM)
region (mesoblastic phase), then to the fetal liver
(hepatic phase) and finally resides in the bone
marrow (medullary phase)
- Mesoblastic (yolk sac) phase
 These primitive yet transient yolk sac
erythroblast are important in early
embryogenesis to produce hemoglobin
(Gower-1, Gower-2 and Portland) needed for
delivery of oxygen to rapidly developing
embryonic tissues
 Yolk sac hematopoiesis differs from
hematopoiesis that occurs later in the fetus
and the adult. In that, it occurs
intravascularly, or within developing blood
vessels
- Hepatic phase
 The hepatic phase begins at 5-7 gestational
weeks and is characterized by recognizable
clusters of developing erythroblasts,
granulocytes and monocytes
 The developing erythroblast in this phase
signal the beginning of definitive
erythropoiesis with a decline in the primitive
hematopoiesis in the yolk sac
 Lymphoid cells begin to appear in this phase,
hemopoiesis occurs extravascularly with the
liver remaining the major site of
hematopoiesis during the second trimester of
fetal life
 Production of megakaryocytes also begins
during this phase. The spleen gradually
decreases granulocytic production and
involves itself solely in hematopoiesis
 Fetal hemoglobin (HgF) is the predominant
hemoglobin, but detectable levels of adult
hemoglobin (HgA) may be present
- Myeloid/medullary phase
 Prior to the 5th month of fetal development,
hematopoiesis begins in the bone marrow
cavity
 This transition is called medullary
hematopoiesis because it occurs in the
medulla or inner part of the bone
 Hematopoietic activity, especially myeloid
activity, is apparent during this phase and the
M:E ratio gradually approaches 3:1 (adult
levels)
 Measurable levels of erythropoietin (EPO),
granulocyte colony-stimulating factor (G-CSF),
granulocyte-macrophage colony-stimulating
factor (GM-CSF), and hemoglobins F and A can
be detected
 Blood cell production, maturation and death occur in
the organs of the RES
- RES includes bone marrow, spleen, liver, thymus,
lymph nodes
- RES functions in hematopoiesis, phagocytosis and
immune defense
 At birth, the bone marrow is very cellular with mainly
red marrow, indicating very active blood cell
production
- Red marrow is gradually replaced by inactive
yellow marrow composed of fat
- Under physical stress, yellow marrow may revert
to active red marrow
Pediatric and Adult Hematopoiesis
 Bone marrow
- Newborn – 80-90% of bone marrow is active red
marrow
- Young adult (age 20) – 60% of bone marrow is
active. Hematopoiesis is confined to the proximal
ends of large flat bones, pelvis, and sternum
- Older adult (age 55): 40% of bone marrow is
active; 60% is fat
- Cellularity – ratio of marrow cells to fat (red
marrow/yellow marrow) and is described in
adults as
 Normocellular – marrow has 30-70%
hematopoietic cells
 Hypercellular/hyperplastic – marrow has
>70% hematopoietic cells
 Hypocellular/hypoplastic – marrow has
<30% hematopoietic cells
 Aplastic – marrow has few or no
hematopoietic cells
- M:E (myeloid:erythroid ratio) – ratio of
granulocytes and their precursors to nucleated
erythroid precursors. Normal ratio is between 3:1
and 4:1
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  Granulocytes are more numerous because of
their short survival (1-2 days) as compared to
erythrocytes with a 120 day life span
 Lymphocytes and monocytes are excluded
from the M:E ratio
- Stem cell theory – hematopoiesis involves the
production of pluripotent stem cells that develop
into committed progenitor cells (lymphoid or
myeloid) and finally mature blood cells
 Progenitor cells
o Lymphoid – different into either B or T
lymphocytes in response to cytokines,
interleukins, growth factors,
lymphokines, CSFs
o Myeloid – give rise to the multipotential
progenitor CFU-GEMM (colony-forming-
unit-granulocyte-erythrocyte-
macrophage-megakaryocyte), which will
differentiate into committed progenitor
cells and finally mature blood cells in
response to cytokines, interleukins,
colony stimulating factors, growth factors
 Lymphoid tissue
- Primary lymphoid tissue
 Bone marrow – site if pre-B cell
differentiation
 Thymus – site of pre-T cell differentiation
 Ag-independent lymphopoiesis
- Secondary lymphoid tissue
 B and T lymphocytes enter the blood and
populate secondary lymphoid tissue, where
antigen contact occurs
 Includes lymph nodes, spleen, gut-associated
tissue (Peyer’s patches)
 Ag-dependent lymphopoiesis depends on
antigenic stimulation of T and B lymphocytes
Bone marrow
 Bone marrow is one of the largest organs located
within the cavities of the cortical bones
 Two major components
- Red marrow – active marrow consisting of
developing blood cells and their progenitor
- Yellow marrow – inactive marrow, composed
primarily of adipocytes (fat cells) with
undifferentiated mesenchymal cells and
macrophages
 During infancy and early childhood, all the bones in
the body contain primarily of red (active) marrow
- Between 5 and 7 years of age, adipocytes become
more abundant and begin to occupy the spaces in
the long bones previously dominated by active
marrow
 The process of replacing the active marrow by
adipocytes (yellow marrow) during development is
called regression
- Eventually results in restriction of the active
marrow in the adult to the sternum, vertebrae,
scapulae, pelvis, ribs, skull and proximal portion
of the long bones
 Hematopoietically inactive yellow marrow is scattered
throughout the red marrow so that in adults, there is
approximately equal amounts of red and yellow
marrow in these areas
 Yellow marrow is capable of reverting back to active
marrow in cases of increased demand on the bone
marrow, such as excessive blood loss or hemolysis
 Adipocytes are large cells with a single fat vacuole;
role in regulating the volume of the marrow; also
secrete cytokines or growth factors that may stimulate
HSC numbers and homeostasis
 Others that are important in hematopoiesis
include: endothelial cells and reticular adventitial
cells, macrophages and lymphocytes, stromal cells
 Hematopoietic microenvironment or niche, plays
an important role in nurturing and protecting HSCs
and regulating a balance among their quiescence, self-
renewal and differentiation
 Stromal cells form an extracellular matrix in the niche
to promote cell adhesion and regulate HSCs through
complex signaling networks involving cytokines,
adhesion molecules and maintenance proteins
Liver
 The liver serves as the major site of blood cell
production during the second trimester of fetal
development
 The hepatocytes in the liver have many functions:
- Protein synthesis and degredation
- Coagulation factor synthesis
- Carbohydrate and lipid metabolism
- Drug and toxin clearance
- Iron recycling and storage
- Hgb degredation in which bilirubin is conjugated
and transported to the small intestine for eventual
excretion
 Kupffer cells are macrophages that remove senescent
cells & foreign debris from the blood that circulates
through the liver; also secrete mediators that regulate
protein synthesis in the hepatocytes
 Liver pathophysiology
- In porphyrias, hereditary or acquired defects in
the enzyme involved in heme biosynthesis result
in the accumulation of the various intermediary
poryphrins that damage hepatocytes, erythrocyte
precursors and other tissues
- In severe hemolytic anemias, the liver increases
the conjugation of bilirubin and the storage of iron
- The liver sequesters membrane-damaged RBCs
and removes them from the circulation
- It can maintain hematopoietic stem and
progenitor cells to produce various blood cells
(called extramedullary hematopoiesis) as a
response to infectious agents or in pathologic
myelofibrosis
- It is directly affected by storage diseases of the
monocyte/macrophage (Kupffer) cells as a result
of enzyme deficiencies that cause hepatomegaly
with ultimate dysfunction of the liver (Gaucher
dane.
 disease, Nieman-pick disease, Tay-Sach’s
disease)
Spleen
 Largest lymphoid organ and has three types of splenic
tissue
- White pulp – scattered follicles with germinal
centers containing lymphocytes, macrophages and
dendritic cells
- Marginal zone – surrounds the white pulp and
forms a reticular meshwork containing blood
vessels, macrophages, memory B cells and CD4-T
cells
- Red pulp – composed primarily of vascular
sinuses separated by cords of reticular cell
meshwork (cords of Billroth) containing loosely
connected specialized macrophages
 As RBCs pass through the cords of Billroth, there is a
decrease in the flow of blood, which leads to
stagnation and depletion of RBCs glucose supply.
These cells are subject to increased damage and stress
that may lead to their removal from the spleen
 The spleen also serves as a storage site for platelets. In
a healthy individual, approximately 30% of the total
platelet count is sequestered in the spleen
 Two methods of removing senescent or abnormal
red blood cells from the circulation
- Culling – cells are phagocytized with subsequent
degradation of cell organelles
- Pitting – splenic macrophages remove inclusions
or damaged surface membrane from the
circulating RBCs
 Spleen Pathophysiology
- As blood enters the spleen, it may follow one or
two routes
 Slow-transit pathway – through the red
pulp, in which the RBCs have a more difficult
time passing through the tiny openings
created by the interendothelial junctions of
adjacent endothelial cells
o The combination of the slow passage and
the continued RBC maturation creates an
environment that is acidic, hypoglycemic
and hypoxic
o Increased environmental stress on the
RBCs circulating through the spleen leads
to possible hemolysis
 Rapid-transit pathway – blood cells enter
the splenic artery and pass directly to the
sinuses in the red pulp and continue to the
venous system to exit the spleen
 Aggregates of T-lymphocytes surround arteries that
pass through these germinal centers, forming a region
called periarterial lymphatic sheath or PALS
 Interspersed along the periphery of the PALS are
lymphoid nodules containing primarily B lymphocytes.
Activated B lymphocytes are found in the germinal
centers
 When splenomegaly occurs, the spleen becomes
enlarged and is palpable
- This occurs as a result of many conditions such as
chronic leukemias, inherited membrane or
enzyme defects in RBCs, hemoglobinopathy,
thalassemia, malaria and the myeloproliferative
disorders
 Splenectomy may be beneficial in cases of excessive
destruction of RBCs such as autoimmune hemolytic
anemia when treatment with corticosteroids does not
effectively suppress hemolysis or in severe
hereditary spherocytosis
- It may also be indicated in severe refractory
immune thrombocytopenic purpura or in storage
disease/disorders with portal hypertension and
splenomegaly resulting in peripheral cytopenias
- After splenectomy, platelet and leukocyte count
increase transiently
 In sickle cell anemia, repeated splenic infarcts caused
by sickled RBCs trapped in the small-vessel circulation
of the spleen cause tissue damage and necrosis, which
often results in autosplenectomy
 Hypersplenism is an enlargement of the spleen
resulting in some degree of pancytopenia despite the
presence of a hyperactive bone marrow
- The most common cause is congestive
splenomegaly secondary to cirrhosis of the liver
and portal hypertension
- Often causes thrombosis, vascular stenosis, other
vascular deformities such as aneurysm of the
splenic artery and cysts
Lymph nodes
 Lymph nodes can be divided into an outer region
called the cortex and an inner medulla. An outer
capsule forms trabeculae that radiate through the
cortex and provide support for the macrophages and
lymphocytes located in the node
 The lymph is the fluid portion of blood that escapes
into the connective tissue and is characterized by a
low protein concentration and the absence of RBCs
 After antigenic stimulation, the cortical region of some
follicles develops foci of activated B cell proliferation
called germinal centers. Follicles with germinal
centers are called secondary follicles, while those
without are called primary follicles
 Located between the cortex and medulla is a region
called paracortex which contains predominantly T
cells and numerous macrophages
 The medullary cords lie toward the interior of the LN
(consisting of plasma cells and B cells); three
functions
- Site of lymphocyte proliferation from the germinal
centers
- Involved in the initiation of the specific immune
response to foreign antigens
- Filter particulate matter, debris and bacteria
entering the lymph node via the lymph
 Lymph node pathophysiology
- Lymph nodes by their nature, are vulnerable to
the same organisms that circulate through the
tissue
dane.
 - Sometimes, increased numbers of microorganisms condition are failure to thrive, uncontrollable
enter the nodes, overwhelming the macrophages infections, and death in infancy
and causing adenitis (infection of lymph nodes) - Adults with thymic disturbance are not affected
- More serious is the frequent entry into the lymph because they have developed and maintained a
nodes of malignant cells that have been broken pool of T lymphocytes for life
loose from malignant tumors  Medullary hematopoiesis – blood cell production
- These malignant cells may grow and metastasize within the bone marrow. Begins in the 5th month of
to other lymph nodes in the same group gestation and continues throughout life
 Extramedullary hematopoiesis – blood cell
Thymus production outside the bone marrow. Occurs when the
 The thymus tissue originates from endodermal and bone marrow can’t meet body requirements and
mesenchymal tissue and populated initially by mainly in the liver and spleen, with hepatomegaly and
primitive lymphoid cells from the yolk sac and the splenomegaly
liver
 In adults, T cell progenitors migrate to the thymus Theories on Blood Cell Formation
from the bone marrow for further maturation  Monophyletic theory – all blood cells come from one
 The thymus resembles other lymphoid tissue in that origin stem cell, the hemocytoblast rec
the lobules are subdivided into two areas: the  Polyphyletic theory – also known as the dualistic
cortex (a peripheral zone) and the medulla (central theory which suggest different groups of blood cells
zone) originate from different stem cells
- Both areas are populated with the same cellular - RBC, WBC, platelets – hemohistioblast
components—lymphoid cells, mesenchymal cells, - Monocytes, lumphocytes and plasma cells – tissue
reticular cells, epithelial cells, dendritic cells and hemohistioblast
many macrophages—although in different  Growth factors
proportions Committed Growth factor Mature cell
 The function of the cortex seems to be that of a progenitor
“waiting zone” densely populated with progenitor T CFU- Thrombopoietin, Thrombocytes
cells MEG GM-CSF
- When these progenitor T cells migrate from the CFU-GM CFU- GM-CSF, M-CSF, Monocytes
bone marrow and first enter the thymus, they M IL-3
have no identifiable CD4 and CD8 surface markers CFU-GM CFU- GM-CSF, G-CSF, Neutrophils
(double negative), and they locate to the G IL-3
corticomedullary junction CFU-GM CFU- Erythropoietin, Erythroid
- Under the influence of chemokines, cytokines and E GM CSF, IL-3
receptors, these cells move to the cortex and CFU-Eo GM-CSF, IL-3, IL- Eosinophils
express both CD4 and CD8 (double positive) 5
- Subsequently they give rise to mature T cells that CFU-Ba IL-3, IL-4 Basophils
expresss either CD4 or CD8 surface antigen as
they move toward the medulla Precursor cells
- Eventually, the mature T cells leave the thymus to
 Comprise the third marrow compartment
populate specific regions of other lymphoid tissue,
 Each type of unipotent stem cell matures into a blast
such as the T cell-depended areas of the spleen,
form
lymph nodes, and other lymphoid tissues
- Myeloblast
 The lymphoid cells that do not express the appropriate - Megakaryoblatst
antigens and receptors, or are self-reactive, dies in - Pronormoblast
the cortex or medulla as a result of apoptosis and are - Lymphoblast
phagocytized macrophages
 The medulla contains only 15% mature T cells and Principle of Normal Blood Cells Maturation
seems to be holding zone for mature T cells until they (synchronistic maturation)
are needed by the peripheral lymphoid tissues
 Blood cells mature synchronistically when its nucleus
 The thymus also contains other cell types including B and cytoplasm mature simultaneously. If ever one lags
cells, eosinophils, neutrophils, and other myeloid cells behind the other, asynchronistic maturation is taking
 Hardly recognizable in old age due to atrophy place
 The thymus retains the ability to produce new T cells,  Cytoplasmic changes
however, has been shown after irradiation treatment - Loss of basophilia
that may accompany bone marrow transplantation  The cytoplasm of an immature cell is usually
 Thymus pathophysiology blue or basophilic due to its ribonucleic acid
- Nondevelopment of thymus during gestation content
results in the lack of formation of T lymphocytes.  The more mature the cell, the less basophilic
Related manifestations seen in patients with this because of the lesser RNA content
dane.
 - Elaboration of cytoplasmic granules - In erythrocytes, this is characterized by persistent
 In myeloid cells, the cytoplasm contain cytoplasmic basophilia and late
granules hemoglobinization
 These granules contain some enzymes which - Abnormal cytoplasmic inclusion bodies may be
distinguish the myeloid stem cells from other found in the cytoplasm of both erythrocytes and
cells leukocytes, especially in the granulocytes
 Those with affinity to red dye are called
acidophilic or eosinophilic granulocytes General Characteristics of “Blast” Cells
 Those with affinity to the blue dye are called  Size – large cells with high N:C ratio
basophilic granulocytes  Cytoplasm – very dark blue and small amount in
 Those with affinity to both acid and basic dyes comparison to the size of the nucleus. No granule is
are called neutrophilic granulocytes present
- Elaboration of hemoglobin  Nucleus – large in size as compared to the size of the
 This is a special feature of the maturation of cytoplasm
erythrocytes  Chromatin – reddish purple and indicates the
 At first, the immature cell contains no predominance of DNA, is very fine and very smooth,
hemoglobin resembling individual grains of sand or a very fine net
 Gradually the hemoglobin starts to appear as (euchromatin); no clumping of chromatin materials
the cell becomes mature until the most - Nucleoli which are present are pale light blue in
mature cell contains a standard and maximal staining
amount of it  Differences between immature and mature cells
 Mature red cells do not possess a nucleus Immature cells Mature cells
 Nuclear changes Cell is large Cell becomes smaller
- Structure and cytochemistry Nucleoli present Nucleoli absent
 The immature nucleus is round and the Fine and delicate Coarse and clumped
nuclear chromatin is very delicate, fine and chromatin chromatin
linear and is called eurochromatin
Round nucleus Round, lobulatd or
 As the cell matures, chromatin strands segmented nucleus
become increasingly coarse and clumped and
Dark blue cytoplasm (rich Light blue cytoplasm (less
are called heterochromatin
in RNA) RNA)
 There is reduction of nucleoli
High N:C ratio Low N:C ratio
 Non-staining areas in the nucleus of old cells
are called parachromatin
Cytoadhesion Molecules in Hematopoiesis
 The chromatin is considered as the best basis
of the maturity of the cell  Cytoadhesion molecules are required to modulate
many interactions between hematopoietic cells and
- Shape
 As the cell matures, the shape of the nucleus growth factors, stromal cells, endothelium and
changes too, and is especially true in extracellular matrix
granulocytes in which the nucleus divides into  These cell surface molecules influence induction,
segments or lobes on maturation differentiation and function of hematopoietic cells and
 The older the cell, the more segments or lobes often have different functions
the nucleus possess  Immunoglobulin supragene family, integrins, selectin
- Cell size changes (LEC-CAM)
 Reduction in cell size during maturation is a  Many extracellular matrix components interact with
feature of all cell lines except in receptors on hematopoietic cells. These incluse
megakaryocytic line fibronectin, thrombospondin, hyaluronic acid,
 Generally, all cell lines increase in number but hormonectin and heparan sulfate
the individual cell’s size decreases because
they undergo mitosis Development of Cells
 With the exception of the megakaryocytic Morphological features Usual development
cells, all mature blood cells are smaller than General cell size Decreases with maturity
the immature stages Nuclear cytoplasmic ratio Decrease with maturity
Nucleus Usual development
Principle of Abnormal Cell Maturation (asynchronistic Chromatin pattern Becomes more condensed
maturation) Presence of nucleoli Not visible in mature cells
 Pathologic hematopoiesis results in abnormal nuclear Color Progresses from darker
maturation, abnormal cytoplasmic differentiation and blue to lighter blue, blue-
abnormal size. The development may also be gray or pink
asynchronous Granulation Progresses from no
 Abnormal cytoplasmic differentiation granules to nonspecific to

dane.
 specific granules normoblast erythroblast
Vacuoles Increase with age Orthochromic Metarubricyte Polychromatic
normoblast erythrocyte
Control of Hematopoiesis Polychromatic Polychromatic erythrocyte
 The entry of mature blood cells into the intravascular erythrocyte erythrocyte
space relies upon (reticulocyte)
- Multiplication of developing cells Erythrocyte Erythrocyte
- Gradual maturation  Progenitors
- Orderly release of cells from bone marrow - Bust-forming unit: erythroid (BFU-E)
 The vessels of the sinusoid system have no direct - Colony-forming unit: erythroid (CFU-E)
connections within the extravascular space, therefore, - Both committed to the erythroid cell line
some form of stimulus is required to either allow the  Estimates of time spent at each stage is about one
diapedesis of cells through the endothelium or week for the BFU-E to mature to the CFU-E
physical rupture of the wall or to engorge the  Another one week for the CFU-E to become a
extravascular space within mature cells pronormoblast, first morphologically identifiable RBC
precursor
Erythrocyte Production and Destruction  At the CFU-E stage, the cell completed approximately 3-
5 divisions before maturing further
Erythrocyte  It takes another 6-7 days for the precursor to become
 RBC or erythrocytes: classic example of the biologic mature enough to enter the circulation which
principles regarding their specialized functions and approximately is 18-21 days to produce a mature RBC
their structures which has specific functions from the BFU-E
 Function: carry oxygen from the lungs to the tissues  The mature erythrocytes have a life span of 120 days in
through attachment of oxygen to the hemoglobin (the the circulation
cytoplasmic component of RBCs).  In the erythrocytes cell line, there are typically three
- Other function: return carbon dioxide to the lungs and occasionally as many as five divisions with
and buffering the pH (secondary function only) subsequent nuclear and cytoplasmic maturation of the
 Protection: control the development, daughter cells, from a single pronormoblast there are
production, and normal destruction of RBC in 8-32 mature RBCs that usually result
order to avoid interruptions of oxygen
delivery even in the presence of harsh Important Events during Erythropoiesis
conditions such as loss of blood with Stage of Erythropoiesis Important Event
hemorrhage Proerythroblast Synthesis of hemoglobin
 Iron, RBC metabolism, membrane structure, and Early normoblast Nucleoli disappear
hemoglobin constitute to the foundation of the body’s Intermediate normoblast Hemoglobin sarts
response to diminished the oxygen capacity of the appearing
blood called anemia Reticulocyte Reticulum is formed; cell
enters the capillary from
Normoblastic Maturation site of production
 Erythroblasts: nucleated precursors in the bone Mature RBC Reticulum disappears; cell
marrow (Normoblast is the developing nucleated
attains biconcavity
cells with normal appearance)
 Morphologic identification of blood cell is well-stained
 Megaloblasts (erythroblasts): abnormal appearance
peripheral blood film or bone marrow smear with
of the developing nucleated cells in megaloblastic
modified Romanowsky stain: may be Wright or
anemia – large size
Wright-Giemsa stain which is the most commonly
 Three Nomenclature Used in Naming Erythroid used
Precursors
 Stage of maturation: careful examination of the nucleus
- Erythroblast: used primarily in Europe
and cytoplasm
- Normoblastic: used in the US and has an
- Qualities of Greatest Importance:
advantage of being descriptive of the appearance
 Nuclear chromatin pattern
of the cells
 Nuclear diameter
- Rubriblast: it parallels the nomenclature used for
 Nucleus:Cytoplasm ratio
granulocyte development
 Presence or absence of nucleoli
 Cytoplasmic color
Three Erythroid Precursors Nomenclature Systems
 As the RBCs mature, the appearance is affected
Normoblastic Rubriblast Erythroblastic
- The overall diameter of the cell decreases
Pronormoblast Rubriblast Proerythroblast
- The diameter of the nucleus decreases more
Basophilic Prorubricyte Basophilic rapidly than does the size of the cell; as a result,
normoblast eryhtroblast the N:C ratio also decreases
Polychromatic Rubricyte Polychromatic
dane.
 - The nuclear chromatinpattern becomes coarser,
clumped, and condensed
 The nuclear chromatin of the RBCs is
inherently coarser than that of the myeloid
precursors
 It becomes even coarser and more clumped
as the cell matures, developing a raspberry-
like appearance in which the dark-staining
chromatin is distinct from the almost white
appearance of the parachromatin
 Ultimately, the nucleus becomes quire
condensed with no parachromatic evident at
all and the nucleus is said to be pyknotic
- Nucleoli disappear, nucleoli represent areas
where the ribosomes are formed and are seen
early in cell development as cells begin actively
synthesizing proteins
 As RBCs mature, the nucleoli disappear which
precedes the ultimate cessation of protein
synthesis
- The cytoplasm changes from blue/gray-blue to
salmon pink
 Blueness of basophilia – acidic components
attract the basic stain (methylene blue)
o The degree of cytoplasmic basophilia
correlates with the amount of ribosomal
RNA. These organelles decline over the
life of developing RBC and the blueness
fades
 Pinkness called eosinophilia or acidophilia
– due to accumulation of more basic
components attract the acid stain (eosin)
o Eosinophilia of erythrocytes cytoplasm
correlates with the accumulation of Hgb
as the cell matures
o Thus, the cell starts out being active in
protein production on the ribosomes that
make the cytoplasm basophilic,
transitions through a period in which the
red of Hgb begins to mmix with that blue
and ultimately ends with a thoroughly
salmon-pink color when the ribosomes
are gone only the Hgb remains
Three General Important Characteristics of RBCs
o Oxygen transport, removal of metabolic wastes
o Loss of nucleus is required for function
o Normal life span is 120 days
Erythrocyte Maturation
 Pronormoblast (Rubriblast)
- Earlist TBC
- N:C ratio of 8:1
- 1-3 nucleoli, nucleous has purple red chromatin
- Chromatin: fine clumps
- Deep or dark blue cytoplasm with no granules
 Basophilic normoblast (Prorubricyte)
- N:C ratio of 6:1
- Centrally located nucleus with 0-1 nucleoli
- Chromatin begins to condense, deep purple red
- Cytoplasm is deeper blue but intensely basophilic
RNA
 Polychromatophilic Normoblast (Rubricyte)
- Size: up to 12um with an N:C ratio from 4:1 to 1:1
- Eccentric nucleus with no nucleoli
- Chromatin shows significant clumping
- Increase Hgb
- Cytoplasm: pink and blue – murky gray-blue
 OrthochromicNormoblast (Metarubricyte)
- Size up to 10um with an N:C ratio of 0.5:1
- Eccentric nucleus with small, fully condensed
(pyknotic) nucleus, no nucleoli
- Pale blue to salmon cytoplasm
- Hgb synthesis decreases
 Reticulocyte (salmon-pink)
- Size: up to 10um
- Contains no nucleus with mitochondria and
ribosomes
- Last stage to synthesize Hgb
- Last stage in bone marrow before release to the
blood
- Reference ranges: 0.5-1.5% (adults); 2.5-6.5%
(newborns)
- Supravital stain – enumeration of reticulocytes
- Reticulocyte count: one of the best indicators of
bone marrow function
- Stress reticulocytes: young cells released from
bone marrow after older reticulocytes have been
released – a response to increased need
- Hgb continuous to be produced by the
reticulocytes for approximately 24 hours after
exiting the bone marrow
 Mature erythrocytes
- Size range: 6-8um
- Round, biconcave discocyte
- Salmon with central pallor (clearing in the center)
in Wright stained smear
 Normal cells have a central pallor that is one-
third the diameter of the cell
 Decreased central pallor is seen with
spherocytic disorders
 Central pallor greater than one-third the
diameter of the cell is seen in microcytic
anemias
dane.
 RBC reference ranges in SI units
- Females: 4.0-5.4 x 1012/L (conventional units 4.0-
5.4 x 106/uL)
- Males: 4.5-6.0 x 1012/L (conventional units 4.6-6.0
x 106/uL)
 Erythropoiesis is regulated by erythropoietin
produced in the kidney. Additional regulation
includes:
 Hypoxia due to high altitudes, heart or lung
dysfunction, anemia
 Androgens (male hormones that appear to
enhance the activity of erythropoietin) and
hemolytic anemias (increased erythrocyte
destruction)
Matured Erythrocytes
 Mature RBC has no nucleus, ribosomes, mitochondria
 Small biconcave discs
 Primary component: hemoglobin (1/3 of cell
volume)
 Flexible because of stretchable fibers called spectrin
Developmental Stages of RBCs
Erythropoiesis
Erythrokinetics
 Describing the dynamics of RBC production and
destruction
 Erythron
- The collection of all stages of erythrocytes
throughout the body: the developing precursors in
the bone marrow and the circulating erythrocytes
in the peripheral blood and the vascular spaces
within specific organs such as the spleen
- Is the entirety of erythroid cells in the body
whereas the RBC mass refers only to cells in
circulation
 Hypoxia
- Too little tissue oxygen, stimulus to RBC
production
- Hypoxia of the peripheral blood us detected by the
peritubular fibroblasts of the kidney, which
upregulates transcription of the EPO gene to
increase the production of EPO
 Erythropoietin (EPO)
- Primary hormone that stimulates the production
of erythrocytes
- Able to rescue the CFU-E from apoptosis,
production of antiapoptotic molecules by the
erythroid progenitors
- Shorten the time between the mitosis of the
precursors
- Release reticulocytes from the marrow early
- Reduce the number of mitoses of precursors
- Produced in the kidney in response to hypoxia
through increased rated of transcription of the
gene
- Needs active bone marrow (no deficiency, no
primary bone marrow disease, and no
suppression by drugs or chronic diseases)
- Normal serum level is 20 IU/L
- Elevated in most anemias (up to thousands) but
lowered in anemia of chronic renal failure
 Apoptosis is the programmed cell death, the
mechanisms by which an appropriate normal level of
cell is controlled
- Fas, the death receptor, is expressed by young
normoblasts, and FasL, the ligand expressed by
the older normoblasts. As long as older cells
mature slowly in the marrow, induce death of
unneeded younger cells
Microenvironment of the Bone Marrow
 Survival of RBC precursors in the bone marrow
depends on the adhesive molecules, such as
fibronectin and cytokines that are elaborated by
macrophages and other bone marrow stromal cells
 RBCs are found in erythroid islands, where
erythroblasts are various stages of maturation
surround a macrophage
 As RBC precursors mature, they lose adhesive
molecule receptors to stromal cells and can leave the
bone marrow timing of egress of RBCs
- Egress occurs between adventitial cells but
through pores in the endothelial cells
Erythrocyte Destruction
 Aged RBCs or senescent cells, cannot regenerate
catabolized enzymes because they lack nucleus
(single feature of normal RBCs responsible for limiting
their life span)
 The semipermeable membrane becomes more
permeable to water so the cell swells and becomes
spherocytic and rigid. Those cells that cannot squeeze
dane.
 through the narrow spaces – trapped in the splenic
sieve
 Eryptosis is a nonnucleated version of apoptosis that
is precipitated by oxidative stress, depletion of energy
and other mechanisms that create membrane signals
that stimulate phagocytosis
 Extravascular or macrophage-mediated hemolysis
- Accounts for most normal RBC death
- The signals to macrophages that initiate RBC
ingestion may include
 Binding of autologous IgG to band 3
 Expression of phosphatidylserine on the outer
membrane
 Cation balance changes
 CD47-thrombospondin 1 binding
 Fragmentation or intravascular hemolysis
- Results when mechanical factors rupture the cell
membrane while the cell is in the peripheral
circulation
- This pathway accounts for a minor component of
normal destruction of RBCs

dane.