HEMATOLOGY 1A

Introduction, Chapter 4, Chapter 5

Hematology Composition of Blood
Þ Derived from the Gk word A. Liquid
o “haima” – blood 1. Plasma/Serum
o “logos” – study/ science Ø Noncoagulated à plasma
Ø Coagulated à serum
*male à average 5L of blood B. Solid – Hemocytes
female à average 4.5-5L of blood 1. Erythrocytes
2. Leukocytes
Functions of Blood 3. Thrombocytes (platelets)
Þ Respiratory C. Gaseous
o Blood transfers oxygen from lung to tissue, 1. Oxygen
and clears the tissue from CO2 and vice 2. Carbon dioxide
versa Þ function of blood is for the transfer and removal of
Þ Nutritional Carbon dioxide
o Happens through blood
o Transports different molecules like proteins, Red Blood Cell
glucose and fats Þ have specific life span of 120 days
o Liquid portion à plasma (contains WBC) Þ Anucleated
§ Plasma has the function to o Because hemoglobin will attach to it
transport and nourish the blood Þ Biconcave
cells Þ Appear pink/red because of iron content
Þ Excretory Þ 6-8um
o Moves waste to liver and kidneys o Microcytic à <6-8um
Þ Buffering Action o Macrocytic à 8um
o In the plasma (liquid portion), the wbc is Þ Presence of pallor area
present and it provides the coagulation o Paled area
enzymes (together w the coagulation o Normal size of pallor area:
factors) that maintain the circulation and § Covering 1/3 of the center of RBC
protect blood vessels from trauma o **if it covers 3/4s, then you’re suffering
Þ Regulating Body Temperature from anemia
Þ Transport of Hormones § Spherocytes
o Because blood is the only liquid connective • erythrocytes that are
tissue sphere-shaped rather than
Þ Defense bi-concave disk shaped as
o Because it contains immunocompetent cells normal.
that serves as defense body mechanisms • found in all hemolytic
o Done by the white blood cells/T cells anemias to some degree
o B cells à produce immunoglobulin • abnormal
Þ Polycythemia Vera
Characteristics of Blood o If there is an increase in the number of RBC,
Þ Fluid (in vivoà inside ur body) tendency is your blood becomes
o You have the natural anticoagulants that hyperviscous
maintain the fluidity and composition of o It is a condition NOT a disease
blood
o White portion of blood = plasma (in vivo) White Blood Cell
o Serum (outside blood) Þ Leukocytes
§ component that is neither Þ nucleated
a blood cell nor a clotting factor; it Þ Not really a blood cell
is the blood plasma not including o Because it merely just “hitched” a ride from
the fibrinogens. the stem cells source/bone
§ does not contain white or marrow/lymphoid tissue/thymus to go to its
red blood cells destination, which is the tissue
Þ Red § In the tissue, it is where they
o Due to iron content (heme) encounter the foreign antigen
Þ Slightly Alkaline § In the secondary lymphoid organs,
Þ Specific Gravity à 1.055 it is where the WBC continually
o Determines if the patient has normal matures and differentiates
hemoglobin or not Þ It is the only related grouping of cell families that are
Þ Blood circulates in the heart, veins, arteries and dedicated to protect the body/host from infection or
capillaries injury
 HEMATOLOGY 1A
Introduction, Chapter 4, Chapter 5

Þ Two kinds: Þ Origin: megakaryocytes
o Agranulocytes Þ Shape: round or oval
o Granulocytes Þ Anucleated
Þ Main Function: to protect host from injury or foreign
antigen ***Can’t extract DNA from RBC because has no nucleus
Þ WBC just hitches a ride from source = stem cells ***RNA: include whole blood, including the plasma
Þ WBC also rides lymphoid tissue and thymus ***human cells generally eukaryotes because have true cells
Þ Destination of the WBC is the tissues and secondary Hematopoiesis
lymphoid organs Þ Grk word “haima” à blood
o In these tissues, WBC encounters foreign Þ Grk word “poiesis” à making/creating
antigen Þ The process of the blood cells undergoing formation,
Þ Help neutralize foreign antigen equally development (differentiation) and specialization
Þ Naïve cells (Terminal stage wherein they are mature and perform
o WBC that do not encounter foreign antigens its specific function)
o Die in apoptosis
Þ No specific life span because life extends only if
encounter specific antigens
Þ Chronic leukemia
o extreme increase in WBC
o WBC appears milky (milky maybe because
fats you ate)
o If have high WBC, need to increase diluting
fluid and decrease blood
Þ Leukopenia
o Decreased WBC = less than 5
o A condition
Þ Leukocytosis –
condition in which
there is an
increase in mature
WBC
Þ Leukemia –
disease in which
one has an
increase in
abnormal or
immature WBC
Þ Called “white” because it appears colorless in a
unstained smear

*Plasma – 55% total blood

Cellular Components – 45% total blood

Evacuated Tubes
1. Yellow Cap – blood culture & sensitivity
2. EDTA – for CBC
3. Plain – for blood culture
4. Blue Cap (Sodium Citrate) – protein affinity

Platelet Satellitosis/Platelet Clumping

Þ Effect of EDTA
Þ True blood cells
Þ Thrombocytes
Þ Maintain integrity of blood vessels
Þ Chemical messengers from the moment we suffer
from inflammation = communicate and release
chemicals and call help
Þ Control hemostasis
Þ Normal size: 2-4 um
 HEMATOLOGY 1A
Introduction, Chapter 4, Chapter 5

Hematopoietic System FETAL HEMATOPOIESIS
Þ Primary organs in the Hematopoietic System
o Bone Marrow and Thymus Three Phases of Hematopoietic/Fetal Development
Þ Secondary organs 1. Mesoblastic/Megaloblastic/Yolk Sac
o Liver, Spleen Ø 2-8 weeks life
o Lymph Nodes Ø Occurs intravascularly (within a
Þ Adult Stage: where the organs are participating in the development of blood vessels)
production of blood Ø Doesn’t contribute significantly to
Þ Fetal Stage: where blood cell development happens “definitive hematopoiesis”
Ø Start 19th day of embryological
Origin of Blood Cells: Hematopoietic Stem Cells (HSCs) development
Þ Originate from mesenchymal layer Ø Development is after fertilization
Þ Foundation of adult hematopoietic system Ø Primitive Erythroblast
Þ Where one gets mature blood cells o Produce measurable amount of
Þ Embryo à Produces 1st adult repopulating HSCs Hgb’s of fetal blood, including
Portland, Gower-1, Gower-2
Types of Hematopoetic Stem Cells (HSCs) Ø Angioblast à remaining cells surrounding
1. Totipontential Stem Cells yolk sac from the future blood vessels
Ø Present only in few hours after fertilization Ø Does not contribute significantly to
Ø Most versatile definitive hematopoiesis
o Can develop into any cell type
2. Pluripotential Stem Cells 2. Hepatic
Ø Present in several days after fertilization Ø Begin 4-5 gestational weeks
3. Multipotential Stem Cell Ø Occurs extravascularly
Ø Derived from pluripotential Ø Decline in primitive hematopoiesis
Ø Can be found in adults Ø Start of definitive hematopoiesis
Ø Limited to specific types of cells to form o Development and appearance of
tissues erythroblast, granulocytes,
monocytes, lymphoid cells,
Importance of HSCs megakaryocytes
Þ HSCs are responsible for the continuing daily Ø Liver
production of all mature cell lineages o Center of blood production
o Major site of hematopoiesis
Þ HSCs are capable of self-renewal and multipotential
§ From fetal life to 1st week
differentiation
after birth
Þ Signaling pathways are important control devices of
§ It reaches peak during 3rd
HSCs’ fate
month after birth
o Erythropoietin
Ø Kidney and Spleen
§ Tells Bone Marrow to produce
o Production of B cells
Blood Cells Thymus
Ø
§ A hormone
o Majority of lymphoid cells develop
Þ Have many chemical mediators that may inhibit here
production of blood cells o T cells mature here
o Interleukin Ø T cells and B cells originate from the bone
o Cytokines marrow
Þ Hypoxia – decrease of oxygen Ø Measurable Levels: Hgb F, HbA, HbA2
o Trigger kidney to release erythropoietin Ø In 2-5 months it is not advisable to do
(hormone) reverse typing (detect antibodies)
§ Signals bone marrow to produce Ø Late development of antibodies for babies
blood cells Ø No antibodies and lymphoid cells in the
§ Example of a signaling pathway fetus yet
• Cytokine
• Lymphokine 3. Medullary/Myeloid
o Erythron = total population from immature to Ø Bone marrow I fetal phase
mature Ø Begins on the 5th month of fetal
development
*Chemical mediators – inhibition of production of blood cells Ø Involvement of Bone Marrow
o Red Marrow à Blood Cells
o Yellow Manner à Fats
Ø Major site of Medullary Hematopoiesis:
Medulla à inner part of bone marrow
 HEMATOLOGY 1A
Introduction, Chapter 4, Chapter 5

Ø Myeloid activity is apparent during this
stage of development
Ø Myeloid-to-erythroid ratio approaches to the
adult level of 3:1 by 21 weeks of gestation
Ø End of 6th month, BM becomes the primary
site of hematopoiesis
Ø Measurable Levels: EPO, G-CSF, GM-CSF,
Fetal hbg, HbA2, Adult hgb
Ø Involvement of bone marrow
Ø Mature cells are ready to leave
Ø Mesenchymal cells migrate into core of
bone and differentiate to skeletal and
hematopoietic blood cells

Summary
Þ Primitive Hematopoiesis
o Primary erythroblast
Þ Definitive Hematopoiesis
o Erythroblast, granulocytes, Monocytes,
Megakaryocytes, Lymphoid cells
o Hepatic phase
Þ Medullary Phase
o Measurable levels of:
§ EPO, G-CSF, GM-CSF, fetal hgb,
Hb2A, adult hgb Mature blood cells pass through the basement membrane

ADULT HEMATOPOIESIS
Interacts with the endothelial layer
Main Sites
Ribs
Sternum shell, shoulder blades
Bind to surface via a receptor-mediated process
Vertebrae
Pelvis, proximal ends of femus & humerus

Bone Marrow Cell pass through the pores in the endothelial cytoplasm
Þ Contains development erythroid, myeloid and and are released into the circulation
megakaryocytic & lymphoid cells
Þ There is participation of lymphoid cells
Þ Lymphoid cells further differentiate to T and B cells *Blood exits the marrow via the central longitudinal vein,
Þ Lymphoid cells specifically B cells can develop which runs through the length of the marrow
without further encountering antigen
Þ Have stromal cells which encounter immature
lymphoid cells with help of stimulating factors – they Lymphoid Development
connect and further differentiate to mature B cells Þ Primary Lymphoid Tissue
Þ Antigen dependent – can encounter with antigen o Consists of blood marrow and thymus
Þ Antigen independent – no encounter with antigen (thymus à where T cells and B cells are
derived)
Primary and Secondary Lymphoid Organs Þ Secondary Lymphoid Tissue
1. Bone Marrow o Lymphoid cells become competent
Ø Tissue located within the cavities of cortical o Consists of the spleen, lymph nodes and gut-
bones associated lymphoid tissue
Ø Two kinds: Red Marrow, Yellow Marrow o Lymphoid cells enter and exit the lymphoid
Ø In the bone marrow, most of the cells are tissue 2 times through the blood
precursor cells at various stages of
maturation Theories on Blood Cell Formation
Ø Red cells and B cells must be mature 1. Monophyletic Theory
before leaving Ø Suggests that all blood cells are derived
Ø The mature will leave the bone marrow from a single progenitor stem cell called the
except the T cells because they will pluripotent stem cell
differentiate in the thymus
 HEMATOLOGY 1A
Introduction, Chapter 4, Chapter 5

2. Polyphyletic Theory Hematopoietic Microenvironment/Cells in the Bone Marrow
Ø Suggests that each of the blood cell lineage 1. Endothelial cells regulate the flow of particles
is derived from its own unique stem cell entering and leaving the hematopoietic space
2. Adipocytes à secrete various steroids that influence
Stem Cell Development Can Be Characterized as: erythropoiesis and maintain bone integrity
Þ Capable of self-renewal 3. Macrophage for phagocytosis and secretion of
Þ Give rise to differentiated progeny various cytokines that regulat hematopoiesis
Þ Able to reconstitute the hematopoietic system of a 4. Osteoblst cells are bone forming cells
lethally irradicated host 5. Osteoclast are bone resorbing celcular
6. Reticular cells associated with the formation of
Fate of HSCs reticular fibers
Þ Self-renewal
Þ Differentiation
o The undifferentiated HSC can be Examination of Maturing Blood Cells
differentiated to progenitor cells committed Þ Cluster differentiation: B cell/T cell
to either lymphoid or myeloid lineages Þ **picture of blood smear parts
Þ Apoptosis Þ Nucleus is intact of lymphocyte
Þ Stain: Giemsa/Wright Stain
Three Phases of Hematopoietic Cells
1. Primitive, Multipotential As the Blood Cell Matures:
Ø Most immature group Þ Overall Cell Size
Ø Occurs in mucosa o Decreases as cell matures
Ø Has stem cells arising from mesenchymal Þ Nuclear-Cytoplasmic Ratio
cells o N:C ratio decreases (from 4:1 to 2:1 to 1:1)
Ø Capable of self-renewal and differentiation (3;1 for lymphocytes)
into all blood cell lines o size of N decreases as cell matures, except
Ø Characteristics: E and T (anucleated)
o Multipotent o space occupied by the nucleus in relation to
o Capable of self - renewal the space occupied by the cytoplasm
o Differentiation Þ Nuclear characteristics
Ø Has primitive RBCs from mesodermal cells o Chromatic, nuclear shape, nucleoli
Ø Stem cells o Nuclear shape become very distinctive as
2. Intermediate cell matures
Ø Consists of committed progenitor cells o Lymphocyte à round or oval
destined to develop to distinct cell lines o Monocyte à kidney bean-shaped
Ø Between stem and normal cells o NEB à segmented nucleoli: # nucleoli
3. Mature Cells varies
Ø Most developed with specific functions Þ Cytoplasmic characteristics à stain, granulation
Ø Erythrocytes ,shape, vacuolization, inclusion bodies, quantity of
Ø Monocytes mature once tissue becomes cytoplasm
macrophage
Ø Basophils – most developed cells Granulation
- In size, ranging from very fine to coarse
Hematopoiesis (PPSC) - In color red à azurophilic blue à basophilc orange
Þ Polypotent stem cells – occurs in yolk cells à eoisinophilc
o Capable of making intermediate and - In the amount of granulation per cell
lymphoid stem cells Cell types Maturation time Survival time (in
Þ Lymphoid stem cells: B cell and T cell (in days) days) – stem cell
Þ T helper cells: T helper 1, T helper 2 and T helper 3 to tissue phase
Þ Cytokine production RBCs 3-5 120
Þ B cell Granulocytes 5-6 9-10
o Cant recognize cell antigen Monocytes 5-6days Months to years
o Further differentiate to plasma cell and Lymphocytes Variable Months to years
memory cell Platelets 4-5 10
o Immunoglobulin production
Þ Wright Giemsa stain can differentiate T/ B cells
 HEMATOLOGY 1A
Introduction, Chapter 4, Chapter 5

Erythrocyte: Maturation, Physiology and Life Cycle
3. Rubricyte
Rubriblast NORMOBLAST ERYTHROBLAST Ø Hemoglobin appears for the first time
(Turgeon)
Rubriblast Pronomoblast Proerythroblast Ø No nucleoli are present
Prorubricyte Basophilic Basophilic Ø Cytoplasm à marked gray-blue (pink and
Normoblast Erythroblast blue)
Rubricyte Polychromatophilic Polychromatic
4. Metrarubricyte
Normoblast Erythroblast
Ø Nucleated RBCs
Metarubricyte Orthochromatic/ Orthochromatic
Ø Last w thin nucleus
Orthrochromatophilic/ Erythroblast/
Ø Cytoplasm à orange-red/pink-orange (like
Acidophilic normoblast Pyknotic
Mat. RBC) reflects nearly complete Hgb
erythroblast
production
Reticulocyte Reticulocyte Reticulocyte
Ø Ortho means à the same cells color the
Erythrocyte Erythrocyte Erythrocyte
same as eosin stain

5. Reticulocytes
Ø RNA, mitochondria, less amount of
ribosomes
Ø Phases: BM and circulation 2-3 days BM 1
day circulation
Ø cytoplasm still w/ small amounts of RNA à
polychromasia (mixed pink and blue
staining) polychromatic erythrocyte
Ø With golgi apparatus remnants and residual
mitochondria that allows continued aerobic
metabolism and hemoglobin production
Summary of Stage Morphology – Reticulocyte Ø STAIN NMB or BCB
Þ Reticular apperances caused by remaining RNA
Þ Has mitochondria and ribosomes through anucleated *Diffusely basophilic erythrocyte and polychromatophilic
Þ Supravital stain such as Methylene Blue erythrocyte à terms sometimes associated to the reticulocyte
Þ Next maturational change specifically when seen on the PBS (Peripheral Blood Smear)

*In Wright-Stained Smear, tericulocytes appeared blue, which 6. Mature Erythrocyte

referred to as polychroatophilia or polychromasia Ø Adult RBC contains no mitochondria (no
Þ An elevated reticulocyte count accompanies a protein/Hgb synthesis
shortened RBC survival Ø No nucleus
Þ Reticulocytosis à indicates that the body is trying to Ø Cytoplasm is salmon pink
maintain homeostasis Ø Delivers O2 to the tissue, releases it and
returns to the lungs to be re-oxygenated
Erythropoiesis
1. Rubriblast Mature RBCs
Ø Nucleoli à present __check book__ usually Þ Up to 16 from 1 rubriblast
very faint Þ 8 mature RBCs usually result from a single
Ø Nucleus à round or slightly oval, thin pronormoblast
nuclear membrane Þ A biconcave disc measuring 6-8um in diameter
o Central or slightly eccentric Þ Appear as salmon pink or red staining cell with a
Ø Cytoplasm à small in amount central pallor area that corresponds to the concavity
o Moderately basophilic on the stained blood smear
o Quite blue because of the
concentration of ribosomes Erythropoietin (Major Stimulatory Cytokine of RBC)
2. Prorubricyte Þ Hormone produced by kidney and liver
Ø Hgb production begins (Rodak) Þ Stimulates CFU-E progenitor cells
Ø Nucleoli à 0-1
Þ Erythron à name given to the collection of all stages
Ø Detectable Hgb synthesis occurs
of RBC throughout the body
Ø Length of time à sl. >24 hours
 HEMATOLOGY 1A
Introduction, Chapter 4, Chapter 5

Polycythemia/Polycythemia Vera
Þ Refer to an increased concentration of erythrocytes
(erythrocytosis) in the circulating blood that is above
normal for gender and age
Þ Secondary erythrocytosis: smoking
Þ Relative polycythemia: increase in erythrocytes that
are not related to increased erythropoietin production
Þ Male and females have different levels of hemoglobin
Þ Testosterone, hormone in male, and erythropoietin
participates in RBC production = males have more
RBC than females
Þ Smoking can affect the number of RBCs
Relative Polycythemia
Þ Has no increase in erythropoietin production
 HEMATOLOGY 1A
Introduction, Chapter 4, Chapter 5