HEMATOPOIESIS ▪ Gower-II (2 alpha and 2 epsilon globin chains)

DEFINITION ▪ Portland hemoglobin (2 zeta and 2 gamma globin

chains)
● Continuous, regulated process of renewal,
proliferation, differentiation, and maturation of all HEPATIC PHASE
blood cell lines
● After 3rd month, yolk sac stops producing blood cells and
● Result in the formation, development, and
is replaced by the fetal liver
specialization of all functional blood cells that are
released from the bone marrow into the circulation ● Starts at the 5th – 7th week of gestation (primary site
● Mature blood cells have a limited lifespan (120 days +/- until the 6th month and continue to produce until 1st to
2nd week of life)
20 days for RBCs) and a cell population capable of self-
renewal that sustains the system ● Characterized by recognizable clusters of developing
● A hematopoietic stem cell (HSC) is capable of self- erythroblasts, granulocytes, and monocytes colonizing
the fetal liver, thymus, spleen, placenta, and ultimately
renewal and directed differentiation into all required
the bone marrow
cell lineages
● Development of spleen, thymus, kidney and lymph
STAGES OF HEMATOPOIESIS nodes contributes to the hematopoietic process
(extramedullary hematopoiesis)
● Pre-natal/Primitive hematopoiesis
o Thymus - first organ to be developed and produces
o Mesoblastic phase T cells (T-cell maturation)
o Hepatic phase o Kidney and spleen – B-cell maturation
o Medullary/Myeloid phase – continues to adult
hematopoiesis ● Blood cells formed:
o Definitive erythroblasts
● Adult/Definitive hematopoiesis
o Granulocytes and megakaryocytes: 3rd month
MESOBLASTIC PHASE o Lymphocytes: 4th month
o Monocytes: 5th month
● Also called as the yolk sac phase ● Hemoglobin formed:
● Begins during the 19th - 20th day of gestation in the o Fetal hemoglobin (Hgb F) 🡪 2 alpha and 2 gamma
blood islands of the yolk sac (mesodermal globin chains
extraembryonic layer 🡪 yolk sac) o Adult hemoglobin (Hgb A1and A2) 🡪 detectable
levels
● Hematopoietic activity is confined to erythropoiesis
− Hgb A1 🡪 2 alpha and 2 beta globin chains
● Production of cells lasts until the: 8th to 12th week of
− Hgb A2 🡪 2 alpha and 2 delta globin chains
gestation
▪ Cells produced in the yolk sac: primitive MEDULLARY PHASE/MYELOID PHASE
erythroblasts and angioblasts (form blood vessels)
● Termed medullary hematopoiesis because it occurs in
▪ Primitive erythroblasts
the medulla or inner part of the bone cavity
o important in early embryogenesis to produce
● Primary site of hematopoiesis is the red bone marrow
embryonic hemoglobin (up to the 3rd month
only) starting 24 weeks of gestation (remains throughout life)
o Large, nucleated, and cannot extrude its ● Begins between the 4th and 5th month of fetal
nucleus
development
o Migrates to the Aorta-Gonad Mesonephros
(AGM) region to become definitive ● HSCs and mesenchymal cells migrate into the core of
erythroblasts (produces HSCs) the bone
● Hemoglobin produced (Embryonic hemoglobin): o Mesenchymal cells - type of embryonic tissue which
differentiate into structural elements that support
▪ Gower-I (2 epsilon and 2 zeta globin chains) developing hematopoietic elements
● Myeloid activity is apparent during this stage with a ● Adults:
normal M:E ratio of 3:1 – 4:1 o 40% active marrow
● Detectable cytokines: o Active marrow is restricted to the sternum,
vertebrae, scapulae, pelvis, ribs, skull, and
o Erythropoietin (EPO) proximal portion of the long bones
o Granulocyte colony-
stimulating factor (G-CSF) ● Retrogression 🡪 process
o Granulocyte-macrophage in which the red marrow
colony-stimulating factor is replaced by the yellow
(GM-CSF) marrow
● Hemoglobin formed: ● IMPORANT NOTE:
o Fetal hemoglobin (Hgb F) o
Yellow marrow is
o Adult hemoglobin (Hgb A1and Hgb A2) capable of reverting
back to active marrow in cases of increased
ADULT HEMATOPOIEITIC TISSUE demand on the bone marrow, such as in excessive
blood loss or hemolysis
● In adults, hematopoietic tissue is located in the:
● Marrow cellularity - ratio of the red marrow to the
o Bone marrow - erythroid, myeloid, megakaryocytic,
and lymphoid cells yellow marrow; usually decreases with age
o Lymph nodes, Spleen, Liver, and Thymus o Normocellular: marrow has 30-70% hematopoietic
(extramedullary hematopoiesis) cells
o Hypocellular: marrow has <30% hematopoietic cells
o Hypercellular: marrow has >70% hematopoietic
● Lymphoid tissues cells
▪ Primary lymphoid tissues 🡪 where T and B cells are o Aplastic: marrow has few or no hematopoietic cells
derived ● COLLECTION PROCEDURES:
o Bone marrow and Thymus o Bone marrow aspirate 🡪 obtained by bone marrow
▪ Secondary lymphoid tissues 🡪 lymphoid cells aspiration
o Core biopsy 🡪 obtained by Trephine biopsy
respond to foreign antigens
o lymph nodes, mucosa-associated lymphoid tissue ● COLLECTION SITES:
(MALT), gut-associated lymphoid tissue (GALT) o Posterior superior iliac crest
o Anterior superior iliac crest
BONE MARROW
o Sternum
● Normal bone marrow contains two major components: o Anterior medial surface of the tibia
o Spinous process of the vertebrae, ribs, or other red
o Red marrow 🡪 hematopoietically active marrow
marrow containing bones
(blood cells and precursors)
o Yellow marrow 🡪 hematopoietically inactive ● EQUIPMENTS USED FOR SPECIMEN COLLECTION:
marrow (adipocytes, with undifferentiated
mesenchymal cells and macrophages) − Surgical gloves

● Infancy and early childhood − Shaving equipment

o 80-90% active marrow − Antiseptic and alcohol
o all the bones in the body
pads
contain primarily red
(active) marrow − Drape material
● 5 to 7 years of age: − Local anesthetic injection
o 60% active marrow
− No. 11 scalpel blade
o adipocytes become more
abundant and begin to − Jamshidi biopsy needle or Westerman-Jensen
occupy the spaces in the needle; Snarecoil biopsy needle
long bones previously
dominated by active marrow − 14-to-18-gauge aspiration needle with obturator
● TYPES OF BONE MARROW SPECIMEN:
o Direct Aspirate Smears: wedge-shape smear; avoids
crushing the spicules
o Anticoagulated Aspirate Smears: K3EDTA
o Crush Smears: places additional glass slide directly
over the specimen
o Imprints: closely replicate aspirate morphology
o Concentrate Smears: narrow-bore glass or a plastic
tube such as Wintrobe hematocrit tube
o Histologic Sections: 10% formalin, Zenker glacial
acetic acid, or B5 fixative
o Marrow Smear Dyes: Wright or Wright-Giemsa dyes
● NORMAL MARROW CELLS:
▪ HEMATOPOIETIC CELLS – PHSC, Progenitor cells,
precursor cells
▪ STROMAL CELLS 🡪 originate from mesenchymal cells
that migrate into the central cavity of the bone
− Endothelial cells – Regulates the flow of particles
entering and leaving hematopoietic space
− Adipocytes – Secretes steroids that influences
erythropoiesis and maintains bone integrity
− Macrophages – Phagocytic cell and secretes
cytokines needed for the regulation
hematopoiesis
− Osteoblasts – Bone forming cells; water-bug or
comet appearance
− Osteoclasts – Bone resorbing cells or destroying
cells
− Reticular cells/Fibroblasts – Supports vascular
sinuses and developing hematopoietic cells
RED MARROW
● Composed of hematopoietic cells arranged in extravascular
● C o r d s a
and are
● supported by trabeculae of spongy bone
● The cords are separated from the lumen of the vascular
sinuses by endothelial and reticular adventitial cells
● Hematopoietic cells develop in specific niches within
the cords
o Erythroblasts - develop in small clusters, and the
more mature forms are located adjacent to the
outer surfaces of the vascular sinuses
o Megakaryocytes - are located adjacent to the walls
of the vascular sinuses, which facilitates the release
of platelets into the lumen of the sinus
o Immature (myeloid) granulocytic cells - are located
deep within the cords
● Niches 🡪 aka hematopoietic microenvironment; plays
an important role in nurturing and protecting HSCs and
regulating their quiescence, self renewal, and
differentiation
HEMATOPOIETIC STEM CELLS (HSC)
● Also called has pluripotential hematopoietic stem cell
(PHSC)
● can differentiate into progenitor cells committed to
either lymphoid or myeloid lineages when stimulated
by cytokines
of
● are capable of self-renewal, are pluripotent, apoptosis,
and may give rise to differentiated progeny
● TYPES:
▪ Non-committed progenitor cells or undifferentiated
HSCs
▪ Committed progenitor cells / Lineage specific
progenitors
 − Common myeloid progenitor/CFU-S/CFU-GEMM 🡪 − suggests that each of the blood cell lineages is
granulocytic, erythrocytic, monocytic, and derived from its own unique stem cell
megakaryocytic lineages
● STEM CELL CYCLE KINETICS
− Common lymphoid progenitor 🡪 T, B, and natural o Bone marrow is capable of producing the following
killer lymphocyte and dendritic lineages per kg body weight:
● Lineage-specific progenitors give rise to − 2.5 billion erythrocytes
morphologically recognizable, lineage-specific
− 2.5 billion platelets
precursor cells
− Precursor cells 🡪 blast forms (ex. myeloblast, − 1 billion granulocytes
o HSCs exist in the marrow in the ratio of 1 per 1000
nucleated blood cells
o Mitotic index
− percentage of cells in mitosis in relation to the total
number of cells
− normal value is 1-2%

− Increased mitotic index implies increased

megakaryoblast, erythroblast) proliferation except in megaloblastic anemia
(prolonged mitosis)
STEM CELL SURFACE
RECEPTORS CYTOKINES AND GROWTH FACTORS
● Used for identification and
● regulate the proliferation differentiation, and
origin of HSCs can be
maturation of hematopoietic precursor cells
determined by
immunophenotypic analysis ● are responsible for stimulation or inhibition of
using flow cytometry production, differentiation, and trafficking of mature
blood cells and their precursors are a diverse group of
soluble proteins that have direct and indirect effects on
hematopoietic cells
● EXAMPLES ARE: Interleukins (ILs), Lymphokines,
Monokines, Interferons, Chemokines, Colony-
stimulating factors (CSFs)
● Positive influence:

− KIT ligand, FLT3

ligand, GM-CSF, IL-1,
IL-3, IL-6, and IL-11
● Negative influence:

STEM CELL THEORY AND KINETICS

● STEM CELL THEORY - Describes the origin of

hematopoietic progenitor cells
o Monophyletic theory
− suggests that all blood cells cells are derived from a
single progenitor stem cell called a pluripotent
hematopoietic stem cell (PHSC)
o Polyphyletic theory
 − Transforming growth factor-b, tumor necrosis
factor-a, and interferons

LINEAGE SPECIFIC HEMATOPOIESIS

● Erythropoiesis

● Leukopoiesis 🡪 Myelopoiesis, Lymphopoiesis

● Megakaryopoiesis