Module 01 Hematopeisis

MODULE 01 | MTY 1211 HEMATOLOGY- LEC
FAR EASTERN UNIVERSITY

INSTITUTE OF ARTS AND SCIENCES
DEPARTMENT OF MEDICAL TECHNOLOGY
A.Y 2021-2022 | 1st SEMESTER
BATCH 2023
HEMATOPOIETIC DEVELOPMENT
• Also known as reticuloendothelial system. • Hematopoiesis in the AGM region and the yolk
sac disappear during this stage.
HEMATOPOIESIS • Spleen, kidney, thymus, lymph nodes
• It is a continuous, regulated process of blood cell contribute to the hematopoietic process during
production. this phase.
Including: • FETAL HEMOGLOBIN
• Cell Renewal
Hb F (2 alpha & 2 gamma)
• Proliferation
• Fetal hemoglobin is produced during 4th
• Differentiation
• Maturation month of gestation.
• BLOOD CELLS FORMED:

HEMATOPOIETIC TISSUE - Erythrocytes
• Organs that function at some point in - Granulocytes
hematopoiesis include the liver, spleen, lymph - Megakaryocytes
nodes, thymus, and bone marrow. - Lymphocytes
- Monocytes
STAGES OF HEMATOPOIETIC TISSUE • THYMUS
• Mesoblastic - The first fully developed organ in the fetus
• Hepatic becomes the major sites of T-cell
• Medullary or Myeloid stage production.
• KIDNEY & SPLEEN
HEMATOPOIESIS
- Produces B-cells.
• MEDULLARY
MESOBLASTIC STAGE
- Production of hematopoietic cells in the
• SITE OF HEMATOPOIESIS:
bone marrow.
• YOLK SAC of the human embryo (19th of
• EXTRAMEDULLARY
embryonic development) > AORTA-GONAD
- Production of hematopoietic cells in the
MESONEPHROS (AGM) region
liver and spleen.
• BLOOD CELLS FORMED:
• MYELOID METAPLASIA
• Primitive nucleated erythroblasts- first
- Myeloid cells being produced outside the
month of embryonic life.
bone marrow.
EMBRYONIC HEMOGLOBINS
MYELOID STAGE
• Gower I (2 zeta & 2 epsilon)
• SITE OF HEMATOPOIESIS
• Gower II (2 alpha & 2 epsilon)
- BONE MARROW (5th month of gestation
• Gower III (2 zeta & 2 gamma)
and continues throughout life)
*Not usually detectable after third month of • Hematopoiesis in the liver begins to slowly
gestation. diminish by the last trimester.
• MEASURABLE LEVELS
HEPATIC STAGE 1. Erythropoietin
- hormone responsible for production of
• SITE OF HEMATOPOIESIS: erythrocytes in the bone marrow
• FETAL LIVER (At 5th to 7th gestational (synthesized in kidney).
weeks) - Stimulated by hypoxia/ decrease in O2
Page | 1 MIGUEL, J.R

BATCH 2023
HEMATOPOIESIS
2. G-CSF- committed progenitor cells BONE MARROW
3. GM-CSF- committed progenitor cells
4. Hemoglobin F and A- committed progenitor • NEWBORN: 80%-90% of bone marrow is
cells active.
• ADULT HEMOGLOBINS • YOUNG ADULT: 60% of the bone marrow is
1. Hb-A1 (2 Alpha & 2 beta) active. Hematopoiesis is confined to the
2. Hb- A2 (2 alpha & 2 delta) proximal ends of large flat bones, pelvis and
sternum.
GLOBIN SYNTHESIS • OLDER ADULT: 40% of bone marrow is active.
CELLULARITY
• Normocellular: marrow has 30% to 70%

hematopoietic cells.
• Hypocellular: marrow has <30%
• Hypercellular: marrow has >70%
• Aplastic: marrow has few or no
M:E RATIO
• 2:1 to 4:1 (normal)

TERMINOLOGIES
• Life Span: 120 days for RBC
PITTING ASPIRATE
• Process that removes inclusions while leaving • Obtained by bone marrow aspiration.
the rest of the red cell intact. CORE BIOPSY
CULLING • Obtained by trephine biopsy
• Process of removing aged or abnormal red COLLECTION SITES:
blood cells from the circulation.
• Posterior superior iliac crest
RBC- 120 days of life span • Anterior superior iliac crest
SUBSTANCE NEEDED: • Sternum
• Anterior medial surface of tibia
1. Iron- Must be in the ferrous state (Fe 2+) • Spinous process of the vertebrae, ribs, or
to transport oxygen other red-barrow containing bones.
2. Amino acid- Globin-chain synthesis
3. Folic acid/ Vitamin B12: DNA WHY?
- The iliac crest is preferred for safety
4. Others- Erythropoietin, Vitamin B6
(Pyridoxine), Trace minerals reasons because no major blood vessels or
organs are located close to this area.
Typically, sampling from the iliac crest also
results in less pain to the patient and is

BATCH 2023
conveniently located for the practitioner NORMAL MARROW CELLS

performing the procedure. • Megakaryocytes: largest bone marrow cell
• Metamyelocyte: most common cell in a
SPECIMEN COLLECTION normal marrow
• Surgical gloves • Macrophages: (alpha naphthyl acetate or
• Shaving equipment alpha naphthyl butyrate) esterase
• Antiseptic and alcohol pads • Mast cells: (+) chloroacetate esterase
• Drape material • Osteoblasts: Synthesized new bone matrix;
• Local anesthetic injection water bug/ comet appearance.
• No. 11 scalpel blade • Osteoclasts: large and multinucleated
• Jamshidi needle or Westerman-Jensen needle;
Snarecoil biopsy needle. MINOR FUNCTION
• 14-to-18-gauge aspiration needle with • Antigen processing of cellular and humoral
obturator. immunity.
BONE MARROW SPECIMEN MAJOR FUNCTION

• Direct Aspirate Smears: wedge-shape smear; • Production and proliferation of blood cells
avoids crushing the spicules. (hematopoiesis)
• Anticoagulated Aspirate Smears: K3 EDTA
• Crush Smears: Places additional glass slide THREE MAJOR CELL TYPES:
directly over the specimen. • Stem cells
• Imprints: closely replicate aspirate • Progenitor cells
morphology • Precursor cells
• Concentrate Smears: Narrow-bore glass or a
plastic tube such as Wintrobe hematocrit tube.
• Histologic Smears: 10% formaline, Zenker
glacial acetic acid, or B5 fixative.
• Marrow Smear Dyes: Wright or Wright-
Giemsa dyes
CYTOCHEMICAL DYES
CYTOCHEMICAL DYE APPLICATION

Myeloperoxidase (MPO) Detects myelocytic cells by staining
cytoplasmic granular contents.
Sudan Black B (SBB) Detects myelocytic cells by staining
cytoplasmic granular contents.
Periodic Acid-Schiff (PAS) Detects lymphocytic cells and
certain abnormal erythrocytic cells
by staining of cytoplasmic
glycogen.
Esterases Distinguish myelocytic from
monocytic maturation stages
(several esterase substrates)
Tartrate-resistant acid Detects tartrate-resistant acid
phosphatase (TRAP) phosphatase granules in hairy cell
leukemia.

BATCH 2023
STEM CELLS - CFU-MEG

• PLURIPOTENTIAL or MULTIPOTENTIAL cells, - CFU-GM
retain the ability to differentiate into any cell
lines. PRECURSOR CELLS
• Often referred to as colony-forming units-
• Blast forms
spleen (CFU-S)
• Comprise the third marrow compartment
• EXAMPLES:
CFU-S: Differentiates in either of two pathways
- Myeloblast
CD2, CD3 Lymphoid, pan T cells - Megakaryoblast
CD4 Helper/ Inducer T cells - Erythroblast
CD8 Suppressor/ Cytotoxic
T cells GENERAL MATURAIONAL CHARACTERISTICS OF
CD13 Pan myeloid CELLS
CD11c, CD14 Monocytes
CD19, CD20 Lymphoid, pan B cells CHARACTERISTICS OF CELLS
CD33 Pan myeloid cells
SIZE: Decreases
CD34 Stem cell marker
N:C RATIO: Decreases
(lymphoid and myeloid
precursor)
NUCLEUS
CD16, CD56 NK cells
Chromatin pattern: more condensed
Presence of nucleoli: Not Visible
PROGENITOR CELLS
CYTOPLASM
ABBREVIATION CELL LINE Color: Darker blue to lighter blue, blue-gray or
CFU-GEMM Granulocyte, pink
erythrocyte, Granulation: No granules to non-specific (primary
megakaryocyte, granules) to specific (secondary granules)
monocyte Vacuoles: Increases with age
CFU-E Erythrocyte
CFU-Meg Megakaryocyte
CFU-M Monocyte
CFU-GM Granulocyte, monocyte
CFU-BASO Myeloid to basophil
CFU-EU Myeloid to eosinophil
CFU-G Myeloid to neutrophil
CFU-pre-T T lymphocyte
CFU-pre-B B lymphocyte
BONE MARROW
PROGENITOR (COMMITED CELLS)

• Unipotential stem cells
• Differentiated into only one cell line.
• INCLUDES:
- BFU-E
- CFU-E
BATCH 2023
ERYTHROPOIESIS
• BONE MARROW: 1%
Normoblastic Rubriblastic Erythroblastic • PERIPHERAL BLOOD: 0%
Pronormoblast Rubriblast Proerythroblast
Basophilic Prorubricyte Basophilic
normoblast erythroblast
Polychromatic Rubricyte Polychromatic
(polychromatophilic) (polychromatophilic)
normoblast Erythroblast
Orthochromic Metarubricyte Orthochromic
normoblast erythroblast
Polychromatic Polychromatic Polychromatic
(polychromatophilic) (polychromatophilic) (polychromatophilic)
erythrocyte erythrocyte erythrocyte
Erythrocyte Erythrocyte Erythrocyte
*Polychromatic erythrocytes are called
reticulocytes when observes with vital fluids.
PRORUBRICYTE
ERYTHROCYTE MATURATION
• Last stage with nucleolus
SIZE: 10 to 15 um
NUCLEUS: round to slightly oval
• NUCLEOLI: 0 to 1
• CHROMATIN: slightly condensed
CYTOPLASM: dark blue
N:C RATIO: 6:1
REFERENCE INTERVAL
• BONE MARROW: 1% to 4%
• PERIPHERAL BLOOD: 0%
ERYTHROPOIESIS
RUBRIBLAST
• Largest in erythroid lineage
SIZE: 12 to 20 um
NUCLEUS: round to slightly oval
• CHROMATIN: fine
CYTOPLASM: dark blue; may have prominent golgi
N:C RATIO: 8:1
REFERENCE INTERVAL
BATCH 2023
RUBRICYTE
• First stage of hemoglobin synthesis
• Last stage that undergoes mitosis
SIZE: 10 to 12 um
NUCLEUS: round
• NUCLEOLI: none
• CHROMATIN: quite condensed
CYTOPLASM: gray-blue as a result of
hemoglobinization
N:C RATIO: 4:1
REFERENCE INTERVAL
RETICULOCYTE
• Peripheral blood (first)
• Last stage of hemoglobin synthesis
SIZE: 8 to 8.5 um
NUCLEUS: absent
• NUCLEOLI: N/A
• CHROMATIN: N/A
CYTOPLASM: color is slightly more blue/ purple
than the mature erythrocyte
N:C RATIO: N/A
REFERENCE INTERVAL
• BONE MARROW: 1%
• PERIPHERAL BLOOD: 0.5% to 2%
METARUBRICYTE
• Most common cell in bone marrow
• Last stage with nucleus
SIZE: 8 to 10 um
NUCLEUS: round
• NUCLEOLI: none
• CHROMATIN: fully condensed
CYTOPLASM: more pink or salmon than blue
N:C RATIO: 0.5 : 1
REFERENCE INTERVAL

BATCH 2023
ERYTHROCYTE
SIZE: 7 to 8 um
NUCLEUS: absent
• NUCLEOLI: N/A
• CHROMATIN: N/A
CYTOPLASM: salmon with central pallor of about
one-third of the diameter of the cell
N:C RATIO: N/A
REFERENCE INTERVAL
• BONE MARROW: N/A
• PERIPHERAL BLOOD: predominant cell
type
PROMYELOCYTE
• First stage of primary granules/ azurophilic
granules/ non-specific granules
SIZE: 14 to 24 um
NUCLEUS: round and oval
• CHROMATIN: fine, but slightly coarser than
myeloblast
CYTOPLASM: basophilic
• PRIMARY GRANULES: more than 20; red
to purple or burgundy
• SECONDARY GRANULES: none
N:C RATIO: 3:1
GRANULOPOIESIS REFERENCE INTERVAL
MYELOBLAST • PERIPHERAL BLOOD: 0%
• Earliest granulocytic precursor
SIZE: 15 to 20 um
• CHROMATIN: fine
CYTOPLASM: moderate basophilia
• GRANULES: absent or up to 20
N:C RATIO: 4:1
REFERENCE INTERVAL

BATCH 2023
MYELOCYTE
• Secondary and specific
SIZE: 12 to 18 um
• NUCLEOLI: usually not visible; slightly eccentric
• CHROMATIN: coarse and more condensed than
promyelocyte
CYTOPLASM: slightly basophilic to creamed-
colored
• PRIMARY GRANULES: few to moderate
• SECONDARY GRANULES: variable number
N:C RATIO: 2:1
REFERENCE INTERVAL
• PERIPHERAL BLOOD: 0% NEUTROPHILIC BAND
• Precursor to neutrophil
SIZE: 10 to 15 um
NUCLEUS: constricted but no threadlike filament;
indented
• NUCLEOLI: not visible
• CHROMATIN: coarse, clumped
CYTOPLASM: pale pink to colorless
• PRIMARY GRANULES: few
• SECONDARY GRANULES: abundant
N:C RATIO: cytoplasm predominates
REFERENCE INTERVAL
• PERIPHERAL BLOOD: 0% to 5%
METAMYELOCYTE
• Most common in bone marrow and first stage
of indentation
SIZE: 10 to 15 um
NUCLEUS: indented; kidney-bean shaped
• NUCLEOLI: not visible
• CHROMATIN: moderately clumped
CYTOPLASM: pale pink to cream colored to
colorless
• SECONDARY GRANULES: many
N:C RATIO: 1.5 : 1
REFERENCE INTERVAL

BATCH 2023
MATURE GRANULOCYTES
SEGMENTED NEUTROPHIL • Acute destruction represented by pancreatitis,
• No segment- hypo segmentation colitis, myocardial infection, severe hemolysis,
SIZE: 10 to 15 um surgical or traumatic wounds, thermal injury
NUCLEUS: 2 to 5 lobes connected by thin filaments
NUCLEOLI: not visible NEUTROPENIA
• CHROMATIN: coarse, clumped • Chemical toxicity (benzene)
CYTOPLASM: pale pink, cream-colored to colorless • Marrow replacement
• PRIMARY GRANULES: rare • Nutritional deficiencies
• SECONDARY GRANULES: abundant • Cytotoxic drugs (amidopyrine and
N:C RATIO: cytoplasm predominates cephalosporin)
REFERENCE INTERVAL
• BONE MARROW: 3% to 11% EOSINOPHIL
• PERIPHERAL BLOOD: 50% to 70% SIZE: 12 to 17 um
NUCLEUS: 2 to 3 lobes connected by thin filaments
NUCLEOLI: not visible
• CHROMATIN: coarse, clumped
CYTOPLASM: cream colored, may have irregular
borders
• SECONDARY GRANULES: abundant pale orange
to dark orange
N:C RATIO: cytoplasm predominates
REFERENCE INTERVAL
NEUTROPHILS
NEUTROPHILIA
• Bacterial infections
• Appendicitis
• Rheumatoid arthritis
• Parasites (Malaria, Liver flukes)
• Lithium
• Chemicals (Lead, Mercury)
• Drugs (digitalis, phenacetin)
• Corticosteroid
• Myelogenous leukemia
• Venoms (spiders, bees, wasps)
• Actinomyces fungi
• Physiologic neutrophilia

BATCH 2023
EOSINOPHILIA
• Asthma BASOPHILIA
• Hay fever • Immediate hypersensitivity
• Psoriasis • Hypothyroidism
• Eczema • Ulcerative colitis
• Scarlet fever • Estrogen therapy
• Parasitic infections
• Eosinophilic leukemia BASOPENIA
• Acute infections
EOSINOPENIA • Stress
• ACTH administration • Hyperthyroidism
• Increased level of glucocorticoids
BASOPHIL • Chronic urticaria
• Allergic infection LYMPHOPOIESIS

• Contains histamine
• IgE- need LYMPHOBLAST
SIZE: 10 to 14 um • Earliest lymphoid precursor cell

NUCLEUS: 2 to 3 lobes connected by thin filaments SIZE: 10 to 20 um
NUCLEOLI: not visible NUCLEUS: round to oval
• CHROMATIN: coarse, clumped NUCLEOLI: greater than or equal to 1
CYTOPLASM: lavender to colorless CHROMATIN: fine, evenly strained
• PRIMARY GRANULES: rare CYTOPLASM: scant; slightly to moderately
• SECONDARY GRANULES: large, deep purple to basophilic
black; irregularly shaped • GRANULES: none
N:C RATIO: cytoplasm predominates N:C RATIO: 7:1 to 4:1
REFERENCE INTERVAL REFERENCE INTERVAL
• BONE MARROW: less than 1% • BONE MARROW: not defined
• PERIPHERAL BLOOD: 0% to 1% • PERIPHERAL BLOOD: 0%

BATCH 2023
PROLYMPHOCYTE
• Agranulocyte
• Immediate precursor cell of lymphocyte
• First stage of indentation
SIZE: 9 to 18 um
NUCLEUS: round or indented
NUCLEOLI: 0 to 1; usually single
Prominent large nucleolus
CHROMATIN: slightly clumped
CYTOPLASM: light blue
• GRANULES: none
N:C RATIO: 3 to 4:1
REFERENCE INTERVAL
• BONE MARROW: not defined
• PERIPHERAL BLOOD: none
LARGE GRANULAR LYMPHOCYTE
SIZE: 7 to 18 um
NUCLEUS: round or indented may be slightly
indented
NUCLEOLI: occasional
CHROMATIN: condensed, clumped, blocky,
smudged
CYTOPLASM: scant to moderate; sky blue vacuole
may be present
• GRANULES: may be a few azurophilic in larger
lymphocyte
N:C RATIO: 5:1 to 2:1
SMALL LYMPHOCYTE
• Comparable to normocytic RBC
SIZE: 7 to 18 um
NUCLEUS: round or indented may be slightly
indented
NUCLEOLI: occasional
CHROMATIN: condensed, clumped, blocky,
smudged
CYTOPLASM: scant to moderate; sky blue vacuole
may be present
• GRANULES: none in small lymphocyte
N:C RATIO: 5:1 to 2:1
REFERENCE INTERVAL
BATCH 2023
MONOPOIESIS
PLASMA CELL MONOBLAST
• Important in humoral immunity • Earliest precursor for monocyte
• Antibody production SIZE: 12 to 18 um
SIZE: 8 to 20 um NUCLEUS: round to oval; may be irregularly
NUCLEUS: round to oval; eccentric shaped
NUCLEOLI: none NUCLEOLI: 1 to 2; may not be visible
CHROMATIN: coarse CHROMATIN: fine
CYTOPLASM: deeply basophilic, often with CYTOPLASM: light blue to gray
perinuclear clear zone • GRANULES: none
• GRANULES: none N:C RATIO: 4:1
N:C RATIO: 2:1 to 1:1 REFERENCE INTERVAL
REFERENCE INTERVAL • BONE MARROW: not defined
• BONE MARROW: 0% to 1% • PERIPHERAL BLOOD: 0%
PROMONOCYTE
LYMPHOCYTOSIS • First stage of indentation
• Infectious mononucleosis SIZE: 12 to 20 um
• Infectious lymphocytosis NUCLEUS: irregularly shaped; folded; may have
• Cytomegalovirus infection brain-like convolutions
• Bordatella pertussis infection NUCLEOLI: may or may not be visible
• Brucellosis CHROMATIN: fine to lacy
• Toxoplasmosis CYTOPLASM: light blue to gray
• GRANULES: fine azurophilic
LYMPHOPENIA N:C RATIO: 2 to 3:1
• HIV Infection REFERENCE INTERVAL
• BONE MARROW: less than 1%

BATCH 2023
MONOCYTOSIS
• Tuberculosis
• Subacute bacterial endocarditis
• Syphilis
• Protozoal and rickettsial infections
• Brucellosis
• Typhoid
• Gaucher’s disease
• Hodgkin’s disease
• Collagen vascular disease
• Gastrointestinal disease
• Surgical trauma
MONOCYTOPENIA
MONOCYTE • Aplastic anemia
• Found in peripheral blood • Overwhelming infections in
SIZE: 12 to 20 um immunocompromised patients
NUCLEUS: variable; horseshoe shaped, or kidney • Hemodialysis
shaped; often has folds producing brain-like • Epstein-Barr virus infection
convolutions • Steroid therapy
NUCLEOLI: not be visible
CHROMATIN: lacy TISSUE MONOCYTE
CYTOPLASM: blue gray
• GRANULES: may fine granules MACROPHAGE
• VACUOLES: absent to numerous SIZE: 15 to 80 um
N:C RATIO: variable NUCLEUS: eccentric, kidney or egg-shaped,
REFERENCE INTERVAL indented or elongated
• BONE MARROW: 2% NUCLEOLI: 1 to 2
• PERIPHERAL BLOOD: 3% to 11% CHROMATIN: fine, dispersed
CYTOPLASM: abundant with irregular borders;
may contain ingested material
• GRANULES: may coarse azurophilic
• VACUOLES: may be present
N:C RATIO: variable
REFERENCE INTERVAL: rarely, seen in the
peripheral blood during severe sepsis.
MACROPHAGE
• Monocytes- peripheral blood
• Kupffer cells- liver
• Microglial cells- CNS
• Osteoclasts- bone
• Langerhans’ cells- skin
• Alveolar cells- lungs
• Mesangial cells- kidney
• Splenic macrophage- spleen
• Hofbauer cells- placenta
BATCH 2023
PROMEGAKARYOCYTE
• First indentation
SIZE: 15 to 40 um
NUCLEUS: indented
NUCLEOLI: variable
CHROMATIN: condensed
• GRANULES: none
N:C RATIO: 1:2
REFERENCE INTERVAL:
• BONE MARROW: 25%
MEGAKARYOPOIESIS
MEGAKARYOBLAST
• Earliest precursor
SIZE: 10 to 24 um
NUCLEUS: round
NUCLEOLI: 2 to 6
CHROMATIN: homogenous and loosely organized
N:C RATIO: 3:1
REFERENCE INTERVAL:
• BONE MARROW: 20%
MEGAKARYOCYTE
• Largest cell in bone marrow
SIZE: 20 to 90 um
NUCLEUS: 2 to 32 lobes
CYTOPLASM: blue to pink; abundant
N:C RATIO: variable
REFERENCE INTERVAL:
• BONE MARROW: 5 to 10 per 10 objective (100
magnification); 1 to 2 per 50 objective (500
magnification)

BATCH 2023
THROMBOCYTES
PLATELETS
SIZE: 2 to 4 um
NUCLEUS: N/A
CYTOPLASM: light blue to colorless
N:C RATIO: N/A
REFERENCE INTERVAL:
• BONE MARROW: N/A
• PERIPHERAL BLOOD: 7 to 25 per oil
immersion field (x1000 magnification)

Module 01 Hematopeisis

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MODULE 01 | MTY 1211 HEMATOLOGY- LEC

FAR EASTERN UNIVERSITY

• BLOOD CELLS FORMED:

Page | 1 MIGUEL, J.R

• Normocellular: marrow has 30% to 70%

• 2:1 to 4:1 (normal)

Page | 2 MIGUEL, J.R

conveniently located for the practitioner NORMAL MARROW CELLS

BONE MARROW SPECIMEN MAJOR FUNCTION

CYTOCHEMICAL DYE APPLICATION

Page | 3 MIGUEL, J.R

STEM CELLS - CFU-MEG

PROGENITOR (COMMITED CELLS)

Page | 6 MIGUEL, J.R

Page | 7 MIGUEL, J.R

Page | 8 MIGUEL, J.R

Page | 9 MIGUEL, J.R

• Allergic infection LYMPHOPOIESIS

SIZE: 10 to 14 um • Earliest lymphoid precursor cell

Page | 10 MIGUEL, J.R

LARGE GRANULAR LYMPHOCYTE

Page | 12 MIGUEL, J.R

Page | 14 MIGUEL, J.R

Page | 15 MIGUEL, J.R

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