Dr. dr. Susianti, M.Sc.

Dosen Histologi
Fakultas Kedokteran Universitas lampung
 GENERAL FEATURES OF BLOOD

• Consist of the cells and fluid

• flow in a regular indirectional movement within the closed
circulatory system
• Total blood volume + 5 L
• Two parts:

• Blood cells
• Plasma
 10 10
9 9
8 8 Plasma
7 7
6 6
5 5
4 4 Leukocytes
3 3
2 2 Erythrocytes
1 1

Hematocrit tubes with blood, Left : Before centrifugation. Right : After

centrifugation. The red blood cell represent 43% of the blood volume in
the centrifuged tube, between the sediment red blood cells and the
supernatant light - colored plasma is a thin layer of leukocytes; this
layer is called the buffy coat.
Three of blood cells:
» Erythrocytes (Red blood cells)
» Leycocytes (White blood cells)
» Trombocytes (Blood Platelets)
Outside the blood vessels: blood undergoes a
complex reaction
 Coagulation
Studied by spreading a drop on a slide single
layer (Blood smear)
The smears are usually
stained by: Romanovsky-
type dye mixtures containing
eosin and methylene blue
(eg: Wright’s or Giemsa)
 1. Eosinophils
4. Lymphocytes
4

5 5. The blood
2. Basophils 1 platelets
2 3

6 6. Erythrocytes
7
3

3. Neutrophils 7. Monocytes
 COMPOSITION OF PLASMA

• Water: 90% by volume

• Solute:10 %
• Plasma protein (70%)
• Albumin
• Globulin
• Fibrinogen
• Other organic compounds (2.1%)
• Amino acids, glucose, vitamins, lipids
• Inorganic salts (0,9%)
• Sodium, potassium, calsium salt
 ERYTROCYTES
406 x 10 6 /L
1. Normal Structure and Function
• Specialist to transport oxygen
• Cytoplasm: oxygen –binding protein
hemoglobin
•  7-8 m,thick:2.6 m & 0,8 m
• Biconcave shape
• Lack nuclei and cytoplasmic
organelles
• Lifespan: 120 days
2. Abnormalities
• Anisocytosis: Great variation in size
• Macrocytes ( larger than 9 m )
• Microcytes ( smaller than 6 m )
• Nuclear fragments
• Howell-Jolly bodies
• Cabot rings
• Reticulocytes
• RBC which contain a small amount of
residual RER and ribosomes
• Staining: Brilliant cresyl blue
3. Hemoglobin
• Consist of 4 polypeptide subunit
• Iron-containing heme group
• Bind reversible:
• O2  Oxyhemoglobin
• CO2  Carbaminohemoglobin
• Bind irreversible:
• CO  Carboxyhemoglobin
• Postnatal:HbA1,HbA2,HbF
• HbS  Abnormal: Sickle Cell Anemia
 4. Plasmalemma and Stroma

• Hipotonic solution: RBC swell and release their Hb 

Hemolysis
• Stroma : SpectrinAssociated with inner surface of
plasmalemma
• Carbohydrate rich-glycocalyx: Outer surface of plasmalemma
 Blood types: A, B, O groups
 LEUKOCYTES (WBC)

6000 – 10,000/L
Nucleated, larger and less numerous than
erythrocytes
2 groups:
Granulocytes
Agranulocytes
2 granules:
Specific granules (granulocytes)
Azurophilic granules
(granulocytes&agranulocytes)
 1. Agranulocytes

Non segment nuclei

Mononuclear leucocytes
2 cell types:
Lymphocytes (20-25% of WBC)
Monocytes (3-8% of WBC)
LYMPHOCYTES

• Also found outside the blood vessels (lymphatic

organ & connective tissue)
• Never becomes phagocytic
• 2 major functional classes:
• T cells
• B cells
• SIZE:
-  6 – 18 m
- Small (7-8 m), medium, large
 NUCLEUS:
- Spheric & often flatted
- Small lymphocytes: densely
heterochromatic, staining purplish-
blue to black
- Large lymphocytes:larges and less
dense
CYTOPLASM:
- Pale basophilia
- Few purplish azurophilic granules
- Smaller ones: thin ring around
nucleus
- Larger ones: more abundant of rim
- Ribosomes>>, mitochondria <<,
ER <<,small golgi complex <<
MEMORY AND EFFECTOR CELLS
- Antigen  Blast transformation:
Memory and effector cells
B Lymphocytes:
Differentiate into plasma cell
Secrete specific antibody / Imunoglobulin
Humoral Imunity
T Lymphocytes:
Major cell of the cellular imune response
Produce lymphokines
Cytotoxic (killer) cells, Helper T cells, Suppressor T
cells
NULL CELLS:
- Circulating cells, resemble lymphocyte
- Represent circulating stem cells of
lymphocytes/ other blood cell
 MONOCYTES
 Large, similar to large lymphocyte
 Only in the blood  Tissue: mononuclear
phagocyte system
(eg: Kupffer, macrophage)
 SIZE:  12-15 m  20 m
 NUCLEOUS:
– 2-3 nucleoli,ovoid, kidney, horses-shaped,
eccentrically placed,rarely spheric
– Chromatin is less condensed
– Stain reddish-purple
CYTOPLASM:
- Faint blue-gray
- Many small azurophilic granules, no
spesific granules
- >> small mitochondria golgi
apparatus, RER, polyribosomes
2. Granulocytes
• Have segmented nuclei
• Polymorphonuclear leucocytes
• Mature nucleus: 2-7 lobes,connected by thin strand of
nucleoplasm
• Specific granules in their cytoplasm
• Granules: membrane- limited
• Golgi complex>>, mitochondria<<, free
ribosomes<<,RER<<
 NEUTROPHILS
The most abundant leukocytes (60-70% of
WBC)
Also found in loose connective tissue
Terminally differentiated cells
SIZE:  12 m  Up to m
NUCLEUS:
Contain condensed chromatin
Most have 3 lobes
Band, hypersegmented neutrophil
Barr body  drum stick
CYTOPLASM:
With specific granules
Granules: 0.3-0.8 m
Stain salmon pink
Less numerous azurophilic granules are
larger and stain a reddish-purple
Specific granules: alkaline phosphatase
and bacterial cationic proteins called:
phagocytins
Azurophilic granules contain lysosomal
enzymes and peroxidase
 EOSINOPHILS
Only 1-4 % of WBC
Also found in connective tissue
Increase: allergic reactions and parasitic
infection
Decrease: Exogenous corticosteroids
SIZE: Slightly smaller than typical neutrophils,
 9m - 14 m
NUCLEUS:
Contain condensed chromatin
2 lobes with bridges
CYTOPLASM:
Presence of numerous large ( 0.5-1.5 m),
brightly eosinophilic granules (specific
granules) in the cytoplasm
Granules: Peroxidase, acid phosphatse,
cathepsin, ribonuclease and the arginine-
rich major basic protein (MBP)
Internum & eksternum
 BASOPHILS
The least numerous of the circulating
leucocytes (0-1% of WBC)
Major cell type at sites of cutaneous
basophil hypersensitivity
Similar with mast cells
SIZE:  10m - 12 m
NUCLEUS:
Contain highly condensed chromatin
3 lobes twisted into S shape
CYTOPLASM:
Specific granules of basophils, irregular
shapes and vary in size.
Granules stain metachromatically,
appear reddish-violet to nearly black
Specific granules: heparin, histamine
 allergic stimuli
 THROMBOCYTES
 Smallest formed elements
  2 - 5 m
 Bone marrow: megakaryocytes
 150,000 – 300,000 / mm3 (m)
 Lifespan 8 days
 Stain faint blue and variety of purple
granules
 Dense bodies(delta granules), Alpha
granules, Lambda granules
 Marginal bundle
 Important physical role in plugging wounds
and clotting factors
Megakaryocytes and platelet formation
Mature cell types in circulating blood
 HEMATOPOIESIS

• The production of blood cells

• Includes: Proliferation and differentiation hematopoietic stem cells
• Erythropoiesis - Agranulopoiesis
• Leukopoiesis - Lymphopoiesis
• Granulopoiesis - Thrombopoiesis
 GENERAL FEATURES OF
HEMATOPOIESIS

A. HEMATOPOIETIC STEM CELLS

• Undifferentiated mesodermal derivatives
• Pluripotent stem cell type (hemacytoblast)
• Colony-forming units (CFUs)
• Unipotential stem cell type
Differentiation
Differentiationof
ofpluripotential
pluripotentialand
andmultipotential
multipotentialcells
cellsduring
duringhematopoiesis.
hematopoiesis.
 B. Bone marrow

• Medullary tissue
• Primary hematopoietic tissue form the 5th
month of fetal development
• All bones: red marrow  yellow marrow
• Reactivated by increased demand
• Contains: Reticular connective tissue stroma,
hematopoietic cords, vascular sinusoids
 C. Blood Cell LifeSpan

• Blood cell have a lifespan in the circulation

• To keep a constant amount  hematopoiesis must be
continous
 D. Regulation of HEMATOPOIESiS

• Involves specific colony-stimulating factors (CSFs):

• Erythropoietin
• Leukopoietin
• Thrombopoietin

  proliferation and differentiation

 DEVELOPMENT OF
HEMATOPOIETIC TISSUE
A. SITES OF INTRAUTERINE HEMATOPOIESIS

1. Primordial (prehepatic) phase

• The 3th week
• Blood islands in the extraembryonic
mesoderm of the yolk sac
• Endothelium of the primitive blood vessels
• Megaloblastic erythropoiesis
 Primitive erythroblast
2. Hepatosplenothymic phase
• The 2th month
• Hematopoietic stem cells invades liver,
spleen, thymus
a. Liver: granulocytes, platelets, RBC decline
at the 5th month a few weeks after birth
b. Spleen: >> erytrhocytes, << granulocytes
and platelets
Just before birth: lymphopoiesis
c. Thymus: Lymphocytes  T cells
3. Medullolymphatic (definitive phase)
• The 3th month
• Bone marrow & lymphoid tissue
a. Medullary tissue
- Diaphysis of the clavicule (2th-3th month)
- 5th month: bone marrow producing platelets
and all types of blood cells
b. Lymphatic tissue
- Additional lympocytes in the lymphoid
tisues and organs (eg: thymus, lymph
nodes, spleen)
- Before birth: lymph nodes  RBC
 B. SITES OF postnatal HEMATOPOIESIS

• Infancy  hematopoiesis is restricted to

the bone marrow and the lymphoid
tissues
 C. extramedullary HEMATOPOIESIS in disease

• In a dults, erythropoiesis, granulopoiesis

and thrombopoiesis in sites other than the
bone marrow are abnormal
 GENERAL STRUCTURE OF MATURE
HEMATOPOIETIC TISSUE
A. BONE MARROW
• Red bone marrow  active or
hematogenous marrow
• During growth  depleted and replaced
by adipocytes
• In adult: Skull, vertebrae, ribs, sternum,
ilia, proximal epiphyses of some long
bones
• Yellow marrow
RED BONES MARROW
1. Stroma
• Adipocytes (75% red marrow)
• Macrophages
• Reticular connective tissue (adventitial cells/ reticular cells and
the reticular fibers)
2. Hematopoietic cords
• Fill interstices of stroma
• Crowded with overlapping blood cells
• Sinusoids between the cord
3. Bone marrow functions
• Bedside hematopoiesis, destroy old red blood cells
• Macrophages in bone marrow, spleen, liver
breakdown hemoglobin (globin, porphyrin,iron)
 B. LYMPHOID TISSUE AND ORGAN
• Thymus, spleen, lymph nodes and
lymphatic aggregations (tonsils and
peyer’s patches
 ERYTHROPOIESIS
A. GENERAL
• Erythropoiesis in bone marrow
• CFU-Ss  CFU-Es
• Cellular changes during erythroid differentiation:
– Decrease in size
– Condensation of nuclear chromatin
– Decrease in nuclear diameter
– Accumulation of Hb
– Decline in the number of ribosomes
– Ejection of the nucleus
STAGES
STAGESOF
OFDEVELOPMENT
DEVELOPMENTOF
OFERYTHROSITES
ERYTHROSITES AND
ANDGRANULOCYTES
GRANULOCYTES
Proerythroblast
Myeloblast
Polychromatophilic
erythroblast
Promyelocyte
early neutrophilic
myelocyte
Early basophilic
Polychromatophilic
erythroblast myelocyte

Late neutrophilic Early eosinophilic

myelocyte myelocyte

Orthochromatophilic Late basophilic

erythroblast myelocyte
Neutrophilic
Late basophilic
metamyelocyte
myelocyte
Reticulocyte
Eosinophilic
Band cell metamyelocyte

Erythrocyte Mature eosinophil

Mature neutrophil Mature basophile

 b. Stages of erythroid differentiation

1. Proerythroblasts
•  14-19 m
• Contain large, centrally, pale staining nucleous (1 or
2 nucleoli)
• Small amount of cytoplasm basophilia
polyribosomes
2. Basophilic erythroblasts
• Slightly smaller than proerythroblasts
•  13-16 m
• Cytoplasm is more intensely basophilic
• Typically staining a deep royal blue
• Prominent, clear, juxtanuclear cytocenter is often
visible
3. Polychromatophilic erythroblasts
•  12-15 m
• Significant Hb
• Grayish appearance
- Polyribosomes basophilic
- Hb Acidophilic
• Nucleus is smaller than in less mature cells with
more condensed chromatin
4. Normoblasts (orthochromatophilic
erythroblasts)
•  8-10 m
• Cytoplasm acydophilic
• Small eccentrical nuclei with chromatin so
condensed appear black
• Early normoblasts can divide
5. Reticulocytes
• Distinguish from mature erytrocytes when stained
with the supravital dye cresyl blue
• Residual polyribosomes blue staining
• Complete their maturation to become erythrocytes
during the first 24-48 hours  biconcave shaped

Reticulocytes Erythrocytes
 Erythropoiesis

P : Proeritroblast
N1 : Basophilic Erythroblast
N2 : Polychromatophilic
Erythroblast
N3 : Orthochromatophilic
Erythroblast
 LEUKOPOIESIS

• CFU-GM Granulocytes & macrophages

• CFU-G Granulocytes
• CFU-M Macrophages
• CFU-Eo Eosinophilis
 A. granulopoiesis

• Adults: in bone marrow

• CFU-G Neutrophils, basophils, eosinophils
• Structural changes
• Decrease in cell size
• Condensation of nuclear chromatin
• Changes in nuclear shape (flattening
Indentation lobulation)
• Accumulation of cytoplasmic granules
STAGES
STAGESOF
OFDEVELOPMENT
DEVELOPMENTOF
OFERYTHROSITES
ERYTHROSITES AND
ANDGRANULOCYTES
GRANULOCYTES
Proerythroblast
Myeloblast
Polychromatophilic
erythroblast
Promyelocyte
early neutrophilic
myelocyte
Early basophilic
Polychromatophilic
erythroblast myelocyte

Late neutrophilic Early eosinophilic

myelocyte myelocyte

Orthochromatophilic Late basophilic

erythroblast myelocyte
Neutrophilic
Late basophilic
metamyelocyte
myelocyte
Reticulocyte
Eosinophilic
Band cell metamyelocyte

Erythrocyte Mature eosinophil

Mature neutrophil Mature basophile

STAGES OF GRANULOCYTE DIFFERENTIATION

1. Myeloblast
•  15 m
• Morphologically undifferentiated
• Has a large, spheric, euchromatic nucleus with up to
3 nucleoli
• Cytoplas lacks granules, more basophilic than CFU
precursors, less basophilic than proerythroblasts
2. Promyelocytes
•  15-24 m
• Chromatin is more condensed
• Spheric nuclei are often flattened on one side,
contain nucleoli
• Their cytoplasm is more basophilic than
myeloblasts, contains azurophilic granules
3. Myelocytes
•  10-16 m
• Sufficient numbers of specific granules
• 3 immatures granulocyte types:
• Neutrophilic myelocytes
• Eosinophilic myelocytes
• Basophilic myelocytes
• Nuclei: Kidney-shaped, highly condensed
4. Metamyelocytes
•  10-12 m
• 3 types:
• Neutrophilic metamyelocytes
• Eosinophilic metamyelocytes
• Basophilic metamyelocytes
• More specific granules
• Nucleus: deeply indented, condensed chromatin
• Capacity for mitosis is lost
5. Band Cells
•  10-12 m
• 3 band cell types: Neutrophilic bands, eosinophilic
bands, basophilic bands
• Horseshoe shaped nuclei
• 3-5% the circulatory leukocytes
• Final maturation: nuclei chromatin condensation
and lobulation
• Mature granulocytes: neutrophils, eosinophils and
basophils
 Maturation of Granulocytes

M1 : Myelocyte
M2 : Metamyelocyte
M3 : Stab cell
M4 : Mature neutrophil
 B. Agranulopoiesis

• From CFU-Ss
• Morphologic changes during maturation
include a decrease in overall cell diameter,
nuclear diameter and increase in nuclear
heterochromatin content
1. Monocytopoiesis
• CFU derivatives monoblasts
• Promonocyte
• Slightly easier to identify and serves as the
immediate precursor of monocytes
•  15 m
• Large than monocytes
• Pale staining nuclei and basophilic cytoplasm
2. Lymphopoiesis
• Adults: in lymphoid tissues and organ and to lesser
extent in bone marrow
• Lymphoblast: larger than lymphocytes
• Null cells
 THROMBOPOIESIS
• From CFU-megs
• Megakaryoblasts:Inactive megakaryocyte
• Megakaryoblastsmitosis: repeated
DNA replications without cellular or
nuclear divisionendomitosis
• Large cells (100 m) megakaryocytes
• Lobulation of the nucleus and
development of an elaborate demarcation
membran system
 Megakaryoblasts

Erythrocytes&
Megakaryocytes Platelets
 COMPARTMENTS AND THE LIFE CYCLE
OF BLOOD CELL TYPES

a. ERYTHROCYTES
• Functionally discrete organ called the erythron
• 2 compartment:
– Circulating compartment
– Medullary compartment
• Reticulocytes enter circulating 24-48 hours
erythrocytes
• Mature erythrocytes circulate about 120 days
• Iron in the Hb  to the bone marrow by
transferrin
• Iron-free portion of Hb  bilirubin
• Controlled by erythropoietin
• Other factors: iron, intrinsic factors, Vit B12,
folic acid
 B. GRANULOCYTES

4 COMPARTMENT THEY OCCURS:

1. Medullary formation compartment
- Stem cells (granulopoiesis)
- 7 days
2. Medullary reserve compartment
- Granulocytes that have yet to enter circulation
- Neutrophil remain 4 days
3. Circulating compartment
- Mature granulocytes circulating in the blood
 relatively constant
- Most circulate only a few hours
- Stimulated by leukoprotein
4. Marginating compartment
- Cells that have entered the circulation,
attached to the walls of blood vessels
- Confined by diapedesis
- Exchanges between the rest of the
marginating compartment occur continously
- Total spent in circulating compartments:
6-7 hours
 C. LYMPHOCYTES

• Precursor of B and T cells produced in bone

marrow
• Thymus circulation spleen or lymph nodes
 T cells  circulation (lifespan:years)
• Bone marrow  B cells  spleenlymp
nodes & other lymphatic aggregations
(lifespan: 6 weeks)  plasma cells
 d. MONOCYTES
• Bone marrow  circulation: for about 2 days
 walls capillaries and venules  connective
mononuclear phagocyte system
 E. PLATELETS

• Form in the bone marrow

• Influenced by thrombopoietin
• Lifespan: 10 days in circulation
• After clot formation: Unkown
Hematopoiesis